NSERC I2I Grant Proposal - Vaccine Stabilization (2016) - PDF

Institutional Identifier FORM 101 Application for a Grant Date System-ID (for NSERC use only)...

Institutional Identifier FORM 101 Application for a Grant Date System-ID (for NSERC use only) PART I 2016/07/04 386147111 Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN) Filipe Carlos DM Valid 256630 Department Institution that will administer the grant Chemical Engineering McMaster Language of application Time (in hours per month) to be devoted to the proposed X English French research / activity 30 Type of grant applied for For Strategic Projects, indicate the Target Area and the Research Topic; for Strategic Networks indicate the Target Area. Idea to Innovation Title of proposal Validation and Scale Up of Vaccine Stabilization for Ambient Handling Using a Pullulan/Trehalose Formulation Provide a maximum of 10 key words that describe this proposal. Use commas to separate them. vaccine stabilization, in vitro validation, in vivo validation, scale up, optimization, ambient handling Research subject code(s) Area of application code(s) Primary Secondary Primary Secondary 1800 1902 1102 1208 CERTIFICATION/REQUIREMENTS If this proposal involves any of the following, check the box(es) and submit the protocol to the university or college's certification committee. Research involving : Humans Human pluripotent stem cells Animals X Biohazards Indicate if the proposed research takes place outdoors and if you answered YES to a), b) or c) – Appendix A (Form 101) must be completed X NO YES TOTAL AMOUNT REQUESTED FROM NSERC Year 1 Year 2 Year 3 Year 4 Year 5 125,000 0 0 0 0 SIGNATURES (Refer to instructions "What do signatures mean?") It is agreed that the general conditions governing grants as outlined in the NSERC Program Guide for Professors apply to any grant made pursuant to this application and are hereby accepted by the applicant and the applicant's employing institution. Applicant Head of department Applicant's department, institution, tel. and fax nos., and e-mail Chemical Engineering McMaster Dean of faculty Tel.: (905) 5259140 ext. 27278 President of institution [email protected] (or representative) Form 101 (2012 W) The information collected on this form and appendices will be stored Version française disponible in the Personal Information Bank for the appropriate program. PROTECTED B WHEN COMPLETED  2 Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe CO-APPLICANTS I have read the statement "What do signatures on the application mean?" in the accompanying instructions and agree to it. Research/ PIN, family name and initial(s) activity time Organization Signature (hours/month) 501309, Jahanshahi Anbuhi, 80 McMaster S CO-APPLICANTS' ORGANIZATIONS AND/OR SUPPORTING ORGANIZATIONS (if organization different from page 1) It is agreed that the general conditions governing grants as outlined in the NSERC Program Guide for Professors, as well as the statements "What do signatures on the application mean?" and "Summary of proposal for public release" in the accompanying instructions, apply to any grant made pursuant to this application and are hereby accepted by the organization. Family name and given name of signing officer, Signature title of position, and name of organization Sayani, Amyn, A GSK Canada - GlaxoSmithKline Mahoney, James, J. B Advanced Theranostics Inc. Form 101 (2011 W), page 2 of 9 Use additional page(s) if necessary. Version française disponible PROTECTED WHEN COMPLETED  2 - 1 Collaborators (RPP except SNG) Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe Before completing this section, read the instructions for the definition of collaborators in the Eligibility Criteria section of the Program Guide for Professors. COLLABORATORS Research/ PIN, family name and initial(s) activity time Organization / Department (hours/month) Ashkar, A 30 McMaster, Form 101 (2012 W), page 2 - 1 of 9 Use additional page(s) if necessary. Version française disponible PROTECTED B WHEN COMPLETED  3 Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe SUMMARY OF PROPOSAL FOR PUBLIC RELEASE (Use plain language.) This plain language summary will be available to the public if your proposal is funded. Although it is not mandatory, you may choose to include your business telephone number and/or your e-mail address to facilitate contact with the public and the media about your research. Business telephone no. (optional): 001 (905) 5259140 Ext. 27278 E-mail address (optional): [email protected] Developing a technique for safely storing and transporting vaccines is considered the "holy grail" in global health, with huge potential for positive social impact. Current systems rely on "cold" supply chains that are expensive, heavily regulated and require a stable source of energy, to ensure that vaccines are always transported, handled, and stored at controlled temperatures. However, these requirements greatly limit the market access of vaccines, especially to the world's most remote and impoverished regions, while greatly increasing manufacturing costs. Today, our goal is to revolutionize how vaccines are handled. Our team has experimented with a pullulan/trehalose formulation that, once added, encapsulates molecules and protects them from environmental conditions (heat, oxygen). This will allow sensitive reagents - such as vaccines - to be transported and stored at ambient conditions. We have developed a proof-of-concept (in vitro only) using the herpes simplex virus (HSV) as a model system; this performance is a strong proof of concept as HSV, in principle, is more difficult to stabilize relative to other viruses due to its enveloped structure. The goal of this project is to expand the commercial value of our pullulan/trehalose stabilizing technology by a) validating performance in vivo b) assessing technology's ability to stabilize different types of vaccines c) developing scale up and implementation procedures for successful user adoption. All research efforts are directed towards the goal of positioning our technology as an engineering solution and a platform for vaccine stabilization. Our proposed method is easy to use while keeping vaccination procedures the same, does not require extensive capital costs or additional manufacturing steps, and will not require external source of energy during storage or transportation. With cost reduction benefits, combined with the value of solving a global health equity challenge, this technology has great potential for commercial success. Other Language Version of Summary (optional). Form 101 (2012 W), page 3 of 9 PROTECTED B WHEN COMPLETED Version française disponible  4 (RPP) Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe ACTIVITY SCHEDULE (Refer to instructions to see if this section applies to your application. Use additional page(s) if necessary.) Anticipated Anticipated Milestone Description of activities starting date completion date Optimization of Create and evaluate pullulan with variety of MWs range 2016-10-03 2016-12-30 pullulan molecular from 500Da-200kDa. Further evaluate for O2 weights & casting permeability, plasticity, dissolution rate, and viscosity method Formulation Assess mixture compatibility; optimize casting conditions 2016-11-01 2017-04-30 (vaccine trials in and ingredient concentrations (pullulan/trehalose ratios) vitro) to achieve target vaccine (HSV, BCG, influenza, tetanus) stability between room temperature and 45C. Vaccine Trials (in Stabilize HSV with pullulan-based mixture for 35 days. 2016-12-01 2017-06-30 vivo) Inject animal with vaccine, then inject animal with virus two weeks later and monitor survival. Repeat procedure with other vaccines. Alter pullulan-based formulation if necessary. Characterization of Test performance under various temperatures, 2017-03-01 2017-06-15 performance under temperature fluctuations, and humidity levels. varying conditions Scale-up methods Use pipetting system to program and automate liquid 2017-04-01 2017-06-30 for large-scale handling fully. Assess and eliminate potential deviations manufacturing between tablets; monitor potential sources of delays in mass production; fine-tune optimal operating conditions. Develop/source Showcase prototypes as demonstration projects for use in 2017-04-01 2017-08-31 additional partners vaccine-stablization (at room temperature). Reach out to for licensing potential partners for licensing. Source additional technology research collaboartion projects, and test other vaccines for compatibility as requested. Final Report Write and submit final report to NSERC 2017-08-01 2017-10-02 Form 101 (2012 W), page 4 of 9 PROTECTED B WHEN COMPLETED Version française disponible  5 (RPP except SNG, SPG and IRC) Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe See instructions for further details. PROPOSED EXPENDITURES Year 1 Year 2 Year 3 Cash In-kind Cash In-kind Cash In-kind 1) Salaries and benefits a) Students 0 0 0 b) Postdoctoral fellows 0 0 0 c) Technical/professional assistants 80,500 15,000 0 0 0 0 d) 0 0 0 2) Equipment or facility a) Purchase or rental 5,000 0 0 0 0 0 b) Operation and maintenance costs 0 0 0 c) User fees 0 0 0 0 0 0 d) Animal Testing 25,000 0 0 3) Materials and supplies a) 0 0 0 Plastic and Glassware 2,000 0 0 b) Vaccine-Related Items 10,000 0 0 c) 0 0 0 0 0 0 4) Travel a) Conferences 0 0 0 b) Field work 0 0 0 0 0 0 c) Project-related travel 0 0 0 d) 0 0 0 5) Dissemination a) Publication costs 0 0 0 b) 0 0 0 6) Technology transfer activities a) Field trials Patenting Activities 7,500 0 0 b) Prototypes 0 0 0 c) 0 0 0 TOTAL PROPOSED EXPENDITURES 125,000 0 0 Total support from industry 0 0 0 Total support from university Total support from other sources AMOUNT REQUESTED FROM NSERC 125,000 0 0 Form 101 (2011 W), page 5-RPP of 9 PROTECTED WHEN COMPLETED Version française disponible  5 (RPP except SNG, SPG and IRC) Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe See instructions for further details. PROPOSED EXPENDITURES Year 4 Year 5 Cash In-kind Cash In-kind 1) Salaries and benefits a) Students 0 0 b) Postdoctoral fellows 0 0 c) Technical/professional assistants 0 0 0 0 d) 0 0 2) Equipment or facility a) Purchase or rental 0 0 0 0 b) Operation and maintenance costs 0 0 c) User fees 0 0 0 0 d) Animal Testing 0 0 3) Materials and supplies a) 0 0 Plastic and Glassware 0 0 b) Vaccine-Related Items 0 0 c) 0 0 0 0 4) Travel a) Conferences 0 0 b) Field work 0 0 0 0 c) Project-related travel 0 0 d) 0 0 5) Dissemination a) Publication costs 0 0 b) 0 0 6) Technology transfer activities a) Field trials Patenting Activities 0 0 b) Prototypes 0 0 c) 0 0 TOTAL PROPOSED EXPENDITURES 0 0 Total support from industry 0 0 Total support from university Total support from other sources AMOUNT REQUESTED FROM NSERC 0 0 Form 101 (2011 W), page 5-RPP of 9 PROTECTED WHEN COMPLETED Version française disponible  6 Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe Before completing this section, read the instructions for contributions from supporting organizations and consult the Use of Grant Funds section in the NSERC Program Guide for Professors concerning the eligibility of expenditures for the direct costs of research and the regulations governing the use of grant funds , and Guidelines for Evaluating Cost-Sharing Ratios and In-Kind Contributions in University-Industry Collaborations regarding the eligibility of in-kind contributions. Name of supporting organization Advanced Theranostics Inc. CONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS Year 1 Year 2 Year 3 Year 4 Year 5 Cash contributions to direct costs of research (Transfer amounts to page five (5); 0 0 0 0 0 except those for the Ship Time program.) In-kind contributions to direct costs of research Salaries for scientific and technical 1) staff 15,000 0 0 0 0 2) Donation of equipment, software 5,000 0 0 0 0 3) Donation of material 0 0 0 0 0 4) Field work logistics 0 0 0 0 0 5) Provision of services 0 0 0 0 0 6) 0 0 0 0 0 Total of in-kind contributions to direct costs of research 20,000 0 0 0 0 In-kind contributions to indirect costs of research (not leveraged) 1) Use of organization's facilities 0 0 0 0 0 Salaries of managerial and 2) 0 0 0 0 0 administrative staff 3) 0 0 0 0 0 Total of all in-kind contributions 20,000 0 0 0 0 Contribution to postsecondary institution overhead 0 0 0 0 0 Form 101 (2011 W), page 6 of 9 PROTECTED WHEN COMPLETED Version française disponible Filipe, C. Contributions from Supporting Organizations CONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS 1) Salaries for Scientific and Technical Staff $15,000 Advanced Theranostics Inc. has agreed to spend research time and effort integrating our stabilization technology with their proprietary isothermal DNA amplification methods. Research will be geared towards assessing potential of using pullulan-based technology in various disease-detection and diagnostic systems. This is in effort to expand the application of our technology and aligns with our outlined activities. Research time expected has an in-kind value of $15,000. 2) Equipment $5,000 Advanced Theranostics Inc. will be providing their equipment and facilities for research that has a mutual benefit for both parties. We have estimated a modest value of $5,000 for access to such equipment used in intended research.  Advanced Theranostics Incorporated June 27, 2016 Dr. Carlos Filipe Department of Chemical Engineering McMaster University Hamilton, ON L8S 4L8 Re: Letter of support for the Idea to Innovation grant application led by Dr. Filipe Dear Dr. Filipe, I am writing this letter in support of your Idea to Innovation grant application and its associated research. Our mission at Advanced Theranostics Inc. is to develop rapid point-of-care tests for the detection of infectious diseases. I am familiar with your work and credentials through our relationship with McMaster University, and I see potential for utilizing your research within our company. Technological advances that stabilize biological reagents, while integrating seamlessly with our current systems, are highly valued. We are currently investigating the use of isothermal amplification for detecting a number of infectious agents such as influenza and Zika virus in full integrated hand held point-of-care test device. Our goal is to package the reagents in a ready-to-use test device providing the user with a test result in 20 minutes. Having the ability to stabilize reagents for storage and transport, in a cost-effective way, is necessary for ensuring the feasibility of our POC test product. Your work using pullulan as a stabilizing agent has potential for improving both viability and robustness of our diagnostic test devices. Advanced Theranostics will provide an in-kind contribution of personnel time ($15000) and equipment ($5000). This commitment will be used to investigate integration of the stabilizing technology into our POC test device and assess its performance. We will also participate in user validation, and provide you with feedback to further develop and improve the technology. I look forward to collaborating with you and your team on this research, which, if successful, can hold great benefit for bringing molecular point-of-care testing closer to reality. I wish you great success on your Idea to Innovation grant application. Sincerely, James B. Mahony, PhD, F.C.C.M., F.A.A.M. CEO and Founder 1171 Rosethorne Road, Oakville, Ontario L6M1H5 Filipe, C. Budget Justification BUDGET JUSTIFICATION 1) Salaries and Benefits $80,500 Research Assistant: Kevin Pennings $29,500 Mr. Pennings joined Dr. Filipe’s lab since 2013 during his undergraduate studies, and has since greatly developed his research skills. He is currently finishing his Masters degree under Dr. Filipe’s supervision, and is expected to defend his thesis this August. Kevin has already 5 peer-reviewed publications. He has exclusive experience on pullulan, and will be a great asset for further developing the technology. His social, outgoing nature will also assist the team in the organization and planning of an effective bridge between academic research and industry. His salary includes 22% of fringe benefits as per university policy. Kevin will be devoting all his time to this project. Lab Technician: Marianne Chew $51,000 A lab technician at McMaster Immunology Research Center, Marianne Chew will draw upon her experience and provide the necessary support for activities associated with animal testing. She will be hired temporarily, and will dedicate approximately half of her time on this project. Her salary is reflective of her employment terms and also includes 30% of fringe benefits as per university policy. 2) Materials and supplies $37,000 Funds will be used to cover expenses associated with enzymes, viruses, substrates, media, mice etc. necessary for the research. Prior experience is used to estimate a reasonable cost for the research outlined. Items Description Cost Plastic and Glassware Well plates, Eppendorf and Falcon tubes, $2000 pipette tips, flasks, bottles Vaccine-related items HSV, influenza and malaria vaccine, vero $10000 cells, media, buffer Animal Testing Mice, training, handling, animal facility $25000 3) Technology Transfer Activities $15,000 Patenting Activities A total of $7,500 is requested. In addition to McMaster University’s contribution of $7,500, a sum of $15,000 will be used for technology transfer activities. Funds will be used to maintain the filed patent and purchase of non-exclusive license from Auburn University ($7500). Total Budget: $132, 500 Total Request: $125,000 Filipe, C. Relationship to Other Research Support RELATIONSHIP TO OTHER RESEARCH SUPPORT A) Grants Currently Held (1) Title: CREATE Biointerfaces Training Program Funding Source: NSERC Grant amount: $150,000 for the first year and $300,000/year for year 2-6 Status: It is in the 5th year of a 6-year grant Conceptual overlap: No Budgetary overlap: 0% Explanation: The CREATE Biointerfaces Training Program provides unique training to graduate students, undergraduates and postdoctoral fellows in the development, characterization and application of bioactive and stealth biointerfaces for ophthalmic biomaterials and biosensor coatings. This training grant involves 11 labs from McMaster University, University of Toronto and Queen’s University. The PI is Dr. John Brennan and I am a co-applicant. My lab has been allocated with the funding to support partial salary of two graduate students (Xudong Deng, and Hanie Yousefi) to work on (1) developing simple techniques to coat paper to prevent non-specific binding of proteins (Xudong Deng) and (2) creating target reporting surfaces via covalent surface modifications with fluorophores and quenchers (Hanie Yousefi). Therefore there is no conceptual or budgetary overlap with the proposed SPG project. (2) Title: Support for Department Chairs Funding Source: Faculty of Engineering – McMaster University Grant amount: $20,000/y (5 years) Status: It is in the 3th year of a 5-year grant Conceptual overlap: No Budgetary overlap: 0% Explanation: The Dean of Engineering provides yearly support to Department Chairs to pay part of the salary of a PDF. Dr. Sana Jahanshahi Anbuhi is the PDF currently supported through this grant and she is working on starting-up a new company. There is no conceptual or budgetary overlap with the SPG proposal. (3) Title: Advanced Manufacturing of Printable Diagnostic Systems for Bioprocess Control and Personal Medicine Funding Source: Ontario Research Fund and industry partners Grant amount: $1,200,000/y (5 years) Status: It has been awarded and we are entering the first year of the grant Conceptual overlap: Minimal (see explanation below) Budgetary overlap: 0% Explanation: This grant involves the development and use of a modular toolbox of printed components to create novel colorimetric, fluorimetric and electrochemical point-of-care diagnostics using scalable printing technologies. The output from the ORF grant will be a series of sensors to detect a range of clinically important biomarkers for infectious disease, neonatal health, wound healing and neonatal health. This grant involves 6 labs from McMaster University – Dr. John Brennan is the PI and I am a co- applicant (along with Professor Yingfu Li, Alfredo Capretta, Ravi Selvaganapathy and Leyla Soleymani). There are 7 industrial partners (Bruker, Fujifilm-Diamatrix, DNA Genotek, Leveraged Green Energy, Pro-Lab Diagnostics, Proteus, and PE Healthcare) and none of them is a partner for the current SPG proposal. The grant supports a project manager, 6 postdoctoral fellows and 6-8 graduate students per Filipe, C. Relationship to Other Research Support year, and all these researchers will work at McMaster Biointerfaces Institute. We are still in the process of negotiating with the industrial partners for IP agreements and specific project activities. My group will play two roles in the project: (1) developing mechanisms (valves, pumps, etc) to enable microfluidics on paper for biological sample preparation and (2) stabilization of sensing reagents (proteins and their small-molecule substrates) with pullulan. My group is expected to receive the funding to support a PDF and 2 graduate students on these activities. Even though both this project and the proposed CRD project share a common approach to stabilize labile reagents, specific project aims and industrial partners are completely different. Therefore, there is some but minimal conceptual overlap and no budgetary overlap with the CRD proposal. (4) Title: Understanding and Manipulating Interactions of Molecules and Paper Surfaces Focusing on the Development of Low-Cost Tools for Preserving and Monitoring Water Quality Funding Source: NSERC Discovery Grant Program Amount: $35,000/year for 5 years (2013-18) Status: It is in the third year of a 5-year grant Conceptual overlap: No Budgetary overlap: 0% Explanation: The Discovery Grant I currently hold is focused on surface modification of paper substrates to prevent non-specific binding of molecules, more specifically proteins. The other goal in this project is to use high-throughput methods to modify paper surfaces to specifically bind protein targets. This project funds the work of a Ph.D. student (Xudong Deng; partial salary and research cost) and one undergraduate student (Mengsu Chen; salary and research cost). Even though the knowledge made with my discovery grant can be beneficial to the proposed SPG grant, these two grants focus on different materials (protein in the Discovery and functional nucleic acids in the SPG) and different applications (basic research in the Discovery and bacterial detection in the SPG). Therefore, there is minimal conceptual overlap and no budgetary overlap with the SPG proposal. B) Grants Applied (1) Title: DNAPrint: a pathogen-tracking paper sensor platform that integrates functional DNA with nanomaterials Funding Source: NSERC Grant amount: $229,000/year for 3 years (2016-2019) Conceptual overlap: No Budgetary overlap: 0% Explanation: This grant will support 1/3 of a technician, 1 PDF and 3 graduate students and 3 co-op students per year for 3 years to develop new printable DNAzyme based biosensors for detection of Legionella in water towers. (2) Title: Industrial wastewater treatment within the "design space" scientific framework Funding Source: NSERC Grant amount: $102,975 (year 1); $101,475 (year 2) and $100,825 (year for 3) Conceptual overlap: No Budgetary overlap: 0% Explanation: The grant is aimed towards developing new detection and treatment technologies for the processing of industrial wastewater (WW) from industrial and commercial facilities (ICFs) The research program is principally inspired by recent trends in the pharmaceutical industry known as the 'design space' concept, an approach which relies on the development and integration of new technologies whose Filipe, C. Relationship to Other Research Support performance are optimized via high-throughput studies. The proposed research is expected to yield an overall improvement in WW effluent quality which would lead to healthier aquatic ecosystems. Filipe, C. I2I Proposal Section 1: Description of the proposed research 1.1 Synopsis The goal of this project is to develop the commercial value of a simple and low cost method to provide thermal stability to currently available vaccines. Using the herpes simplex virus (HSV-2) as a model system, our group was able to show that HSV-2 can be thermally stabilized and preserved at room temperature for several weeks, by entrapping the virus in a pullulan/trehalose film. Pullulan is a linear polysaccharide that is used as the film forming material in LISTERINE® breadth strips. Pullulan films are impermeable to oxygen and dissolve very rapidly in water. Trehalose is a disaccharide that has been extensively used for preserving biomacromolecules due to its high ability to retain water. We found that in order to effectively preserve HSV-2, both pullulan and trehalose are required in the film forming process. After the vaccine has been entrapped in the films, it can be preserved at room temperature with minimal loss of potency. To reconstitute the vaccine before injection, a saline solution is added to the film and the vaccine is rapidly released into the solution. Current proof-of principle of this was obtained in vitro. The goal moving forward is to verify that the stabilization technology is also effective in vivo and for a variety of different vaccines - this will position the technology as a general platform for thermally stabilizing vaccines. By demonstrating to potential partners the capacity to stabilize a range of vaccine products, in a way that integrates seamlessly with current user and manufacturing processes, we will increase the value of this technology before licensing to industry. Time and effort will be dedicated to develop this thermo stable technology for vaccines that normally require cold chain logistics. In particular, we will use several viral vaccines, including influenza to test their stability and immunogenicity. 1.2 Background We have developed a simple method for storing, handling and transporting vaccines without the need for refrigeration. For the last three years, our lab has been experimenting with adding film-forming pullulan/trehalose mixtures to solutions containing labile molecules such as proteins, enzymes, nucleic acids and easily oxidizable small molecules. Pullulan, the predominant component of the system, is a naturally occurring (and FDA approved), water-soluble polysaccharide produced by the fungus Aureobasidium pullulans. Trehalose is a disaccharide (also FDA approved) extensively used for protecting molecules against drying. We have found that entrapping labile molecules in pullulan/trehalose (P/T) films effectively protects these molecules against thermal degradation and oxidation. Just before the entrapped reagents are to be used, water (or any other buffer) is added to the film, rapidly dissolving the film and releasing the reagents to the solution. Preserving vaccines is a far more challenging goal, but we have obtained proof-of-principle, using herpes simplex virus (HSV-2) as a model system, that proves P/T films can achieve this goal. HSV-2 is an enveloped virus, which, in principle, is more difficult to stabilize than non-enveloped viruses. Figure 1 shows that HSV-2 can be effectively preserved for at least 30 days at room temperature, when entrapped in a P/T film, as compared to keeping the virus in a buffer solution or in a trehalose solution. We realize that there is an initial drop in viral counts during the film-formation period. One of the goals in this project is to minimize this initial loss, using different methods for drying and we will validate the preferred drying approach using multiple vaccines. Figure 1. Plaque forming units (PFU) for HSV-2 as a As mentioned before, we have also function of time for samples kept at room temperature and stored in: buffer (!) buffer; (") a 0.5M trehalose demonstrated the versatility of the P/T film solution; and in pullulan/trehalose (P/T) films (#). technology to stabilize other reagents. We have 1 Filipe, C. I2I Proposal shown, in addition to HSV-2, pullulan-based films can provide long term stability (several months) to many enzymes (such as Taq polymerase for DNA amplification), bacteriophage, proteins and other labile reagents, that have been stored at room temperature (no refrigeration). These results strongly reinforce the versatility of this technology and we want to position ourselves as a stabilizing platform capable of meeting demands of biopharmaceutical manufacturers and their diversified product lines. Developing a technique for safely storing and transporting vaccines is considered a “holy grail” for global health, with huge potential for positive social impact. Addressing this challenge has garnered the interest of many organizations, including the Global Good program (a collaboration between Bill Gates and Intellectual Ventures), which are developing passive vaccine storage devices that can keep reagents cold without an external source of energy. However, at a price of $1,100, the device fails to fulfill the goal of increasing accessibility to the world’s poorest areas. Currently, industry depends on a cold supply chain that ensures products have a means of being transported and stored at cold temperatures. However, this system is highly regulated, costs $100-$105 per square foot more than an ambient facility, and requires a reliable energy source for operation. These requirements highly limit market access. Another common method for handling vaccines is via lyophilisation (freeze-drying). However, this process still requires some products to be stored between 2-8°C, and does not address the challenge in a full manner. Lyophilisation can also cause loss of activity, due to exposure to ice- water interfaces during freezing, pH shifts, and mechanical damage from ice crystals. There is ongoing research for improving the process of lyophilisation, through addition of additives such as trehalose, to protect against damage caused during the process. However, this direction does not eliminate this method’s dependency on a capital and energy intensive process. Research has been directed towards “proprietary formulations” for stabilizing sensitive products. Several emerging companies have been built on such technologies, though none of the identified companies are major players. Furthermore, these emerging companies are only present in some markets, and do not focus on vaccine stabilization (refer to Section 2.2.3 for Competitive Landscape). Relative to these new technologies, we remain positioned as a platform focused on providing solutions for vaccine stabilization. Pullulan-based technology will be branded as the sole, cost-effective and user-friendly method for stabilizing vaccines in the market, meeting needs during storage and transportation. Relative to established systems, this technology will be revolutionary, characterized by lower capital costs and independence from energy requirements– both of which are inherent in cold chain logistics and lyophilisation. The target performance of our technology will be a shelf life of 26 weeks or more, under ambient and elevated temperatures of up to 45°C. This was chosen to perform at least on par with a vaccine stabilized by lyophilisation and a trehalose excipient (a competitive performance determined from literature review). Additional novelty is in the implementation process, which does not require extensive capital costs and/or significant changes to the manufacturing process. Incorporating this technology only requires adding a mixture to a vaccine under refrigerated environments (already present in manufacturing facilities). Using the vaccine in its pullulan-stabilized form also does not require excessive steps or special conditions; addition of saline buffer suffices (the status quo). Both of these characteristics are critical for ensuring user adoption, and as such, research will be focused on fine- tuning the scale-up process, which will be a critical component of technology license. The prevailing market conditions for positioning the technology as a solution provider for vaccines, and subsequently the biopharmaceutical industry, is very favourable. The biotech industry is under a forecasted growth rate at 8% per year, twice the rate of the conventional pharmaceutical industry. Furthermore, majority of the best selling pharmaceutical products by 2018 will be driven by biotechnology. This forecasted growth ensures that our technology will have continued relevance in future years, starting with vaccines (the goal of this project), and later expanding to include other biomolecules as future pipeline products. The ability to handle and store vaccines under ambient conditions for extended periods of time, in a cost-effective way, without relying on energy sources, is needed to supplement growth in the industry. 2 Filipe, C. I2I Proposal This technology does not require complex additional steps to be incorporated into existing manufacturing processes. Additionally, versatility positions the technology as an engineering solution for the industry, not a single product. All these factors indicate that there is untapped possibility for commercial success, ready to be further developed. This project is designed to develop this commercial value. 1.3 Detailed Proposal There are several technical complexities that must be resolved before this technology can be commercialized. Firstly, the viability of this technology as an engineering solution and a platform rests on its ability to meet the demands of a market that has diversified products (i.e. versatile characteristics). Furthermore, in the vaccine market space, the value of a technology is highly dependent on its performance at the in vivo stage (positive in vitro performance only does not suffice). Lastly, customer adoption will be dependent on how well the new technology integrates with existing manufacturing processes during scale-up. Research activities in this project (later outlined) are designed to tackle these core complexities. The I2I project specifically addresses the following key technical objectives necessary for commercial success: (1) assess technology’s performance with multiple vaccines types; (2) characterize performance in vitro and in vivo; (3) establish scale-up and implementation processes for manufacturing. Objective 1: Assess technology’s performance with multiple vaccines types Research activities associated with Objective 1 are tailored to address the need for versatility as a critical characteristic of this technology. Time and effort will be dedicated to stabilizing already available vaccines for influenza, BCG (tuberculosis) and tetanus. It is important to note that in this proposal we will not be developing new vaccines or modifying current vaccines to improve potency, we will only focus on thermal stabilization. The TK- HSV-2 vaccine is a live-attenuated vaccine (virus); the tetanus vaccine is a recombinant bacterial vaccine (protein), whereas the influenza vaccine can be an inactivated virus, live-attenuated or recombinant vaccine. We will use BCG as an example of live attenuated bacterial vaccine. These 4 vaccines are explicitly chosen to provide a comprehensive range of vaccine types to experiment with. Showcasing the technology’s adaptability to different vaccine types will be strategic for demonstrating relevance to industry. This is a critical objective that can influence the success of this project. Failure to adapt to various vaccine types will limit the attractiveness of this technology, and as such, is a critical go/no-go decision point. Referring to the Activity Schedule, this objective will be explored over 7 months in tandem with in vitro and in vivo testing activities. An extension of this objective will also include actively reaching out to source additional partners, showcasing demonstration projects, and turning expressed interest into collaboration or licensing opportunities (also outlined in the Activity Schedule). This step is contingent on the results from earlier activities. The team has access to TK- HSV-2 and BCG through Dr. Ali Ashkar, a collaborator in this proposal, and will get input from GlaxoSmithKline (GSK) in regards to tetanus and influenza vaccines. Go/No-Go: Performance with the four stated vaccine types will determine whether technology can easily adapt to different vaccines, and if this technology is indeed versatile. Risk: There is a technical risk associated with positioning a technology as a “platform” and assessing its future commercial success solely on how well it performs with four vaccines. It might be the case that results obtained provide unusual favourable or unfavourable results, in either case not necessarily reflecting the true potential of the technology. For example, results from these trials might either under or over-value the technology. However, we have specifically chosen vaccines that span different categories. Diversification is meant to mitigate this risk and reduce the chance of all four vaccines performing a certain way, later wrongfully influencing the direction of the project. Furthermore, the TK- HSV-2 vaccine, by nature, is difficult to preserve. Hence, any success with TK- HSV-2 will more likely translate to similar results in other vaccines. 3 Filipe, C. I2I Proposal Objective 2: Characterize performance in vitro and in vivo Both in vitro and in vivo tests will be conducted to characterize the performance of the P/T in thermally stabilizing the vaccines. We will first conduct experiments to confirm that the film preserves the bioactivity of the vaccine in vitro. These will require adjusting the formulation of the P/T films to yield optimal in vitro results for each vaccine at room temperature and 45°C. Casting conditions and ingredients will also be optimized (closely related with some of Objective 3 activities). Preliminary work on the TK- HSV vaccine has already been completed with promising results. However, we need to evaluate the immunogenicity of the vaccines in an animal model in a vaccination challenge platform. The main focus of this stage will be the BCG and influenza vaccines (not investigated before). In vitro testing involves the viability of the vaccine viral titers (for influenza) and bacterial culture (for BCG) using well established protocols in Dr. Ashkar’s lab. The virus will be stabilized for a period of 6 weeks, under temperatures ranging from room temperature (RT) to 45°C, prior to checking the virus’ retained activity. The extent of viral activity is measured via plaque forming units to indicate where the virus successfully infected cells. In vivo testing will involve animal testing conducted in mice. These tests will be done gradually, starting with the TK- HSV-2 vaccine, and with the assistance of a McMaster Immunology Research Center lab technician. The procedure spans approximately 50 days, and requires the vaccine to be stabilized for 35 days with the appropriate pullulan-based formulation (as determined from the in vitro stage) before being administered. Mice will then be vaccinated with the stabilized vaccine and non-stabilized vaccine. Three weeks post-vaccination, mice will be challenged with wild type pathogen to see if vaccination will lead to protection. As positive control, we will use fresh vaccine, which is well known to completely protect mice from subsequent challenge with a lethal does of pathogens. Depending on results, this step might require further tweaking of the P/T based formulation to improve results. The team understands that a feedback loop between in vitro and in vivo results is expected to improve functionality. Further tests will involve increasing storage time to 26 weeks and temperature to 45°C before administering, to accurately characterize extent of stabilization capability. Prior meetings with GSK have indicated that in vivo results are critical for assessing the attractiveness of this technology to industry. In vivo performance is a limiting decision factor, and this stage is suitably a go/no-go decision point as well. The criticality of this stage is well understood by our team. However, proof-of-concept results have been very favourable at the in vitro stage and we are very optimistic that in vivo trials will yield similar results. Regardless, the team is prepared to invest the most time and effort at this stage, to meet our end goal of providing quantitative results to showcase performance and technological viability. Go/No-Go: Results from in vivo testing will provide conclusive data on extent of vaccine stabilization with target storage conditions of 26 weeks and temperatures from RT to 45°C. Risks: There is also a technical risk, though minimal, with injecting the pullulan/trehalose/vaccine mixture into the body. Trehalose has been approved for injection into the human body; pullulan is edible, FDA approved and has been shown to be non-toxic, non-mutageneic, non-carcinogenic and non-immunogenic. It is unlikely that injecting the pullulan/trehalose/vaccine mixture will problematic, but a risk does exist and this is an area that must be explored and tested. Objective 3: Establish scale-up and implementation processes for manufacturing User adoption is influenced by how well Objective 3 is met. Research activities associated with this objective address various aspects of implementation and scale-up processes that are valuable for manufacturing. The first step will evaluate a range of pullulan molecular weights (from 500Da to 200kDa) and casting conditions that optimize and simplify processability (refer to the Activity Schedule. Characteristics to be monitored at this stage are: O2 permeability, plasticity, dissolution rate and viscosity. Trade-offs between characteristics that affect processability (plasticity, viscosity) and those that affect performance (O2 permeability, dissolution rate) will be studied to determine optimal conditions. Performance of P/T substances (using generic reagents and vaccines) will be studied under varying environmental conditions (temperatures above ambient conditions, temperature cycling, humidity). Note that vaccines tested at this stage will be tested in vitro only (this will suffice, as in vivo tests would have 4 Filipe, C. I2I Proposal already confirmed performance). Results will provide manufacturers with further insight on required packaging and storage conditions, as well as performance under varying environmental conditions (expected to change during transportation for example). Different types of drying will explore impact on quality and process times. Methods to be tested include air-drying, spin-drying, and spray drying. These activities are designed to further develop proprietary knowledge that is valuable for manufactures and user adoption. The inability to scale-up and/or integrate with existing manufacturing processes will significantly hinder commercial success. As such, this objective is of great importance. Go/No-Go: Design process will assess the simplicity/complexity involved with integrating technology into existing manufacturing processes, and whether scale-up process meets manufacturing criteria (target throughput, quality control, etc.) for a given client. Risks: The high viscosity of the P/T mixtures may complicate scale-up and operating procedure. However, altering the molecular weight of the pullulan is likely to mitigate this risk. 1.4 Team Expertise Carlos Filipe, PhD. Dr. Filipe is a Professor and Chair of the Department of Chemical Engineering at McMaster University. He has extensive experience in the areas of biochemical engineering, development of ultra-low cost sensors for environmental and health monitoring. His research has been published in top scientific journals, such as the Journal of the America Chemical Society (JACS) and by popular media (CBC, Globe and Mail, FastCompany, etc.). He has received several research and teaching awards and for his last Discovery Grant, he was awarded a Discovery Accelerator Supplement. Dr. Filipe will oversee all aspects of this project. The pullulan/trehalose film technology was developed in Dr. Filipe’s lab. Sana Jahanshahi-Anbuhi, PhD. Dr. Jahanshahi-Anbuhi is a postdoctoral fellow at the Department of Chemical Engineering at McMaster University. She completed her PhD on developing pullulan-based formulations for reagent stabilization. She has 10 publications and four patent applications, two of which cover her work with pullulan. Her extensive experience in pullulan-based research positions her as a lead scientist on this project. She will focus on optimization of pullulan molecular weights, casting methods, and scale-up processes. She has been awarded multiple awards, including the Shell Canada Graduate Research Fellowships Award and A. E. Hamielec Graduate Student Award. She is highly motivated on commercializing this technology. Ali Ashkar, DVM, Ph,D: Dr. Ashkar is a professor at McMaster immunology research Center, Department of Pathology and molecular medicine. He holds a Tire 1 Canada Research Chair (2014- 2021). He has extensive experience in immunology, volcanology, experimental animal models and host- pathogen interactions. He is a collaborator in this project. Vincent Leung, MASc, PhD Candidate. Mr. Leung is a NSERC Vanier CGS Scholar – a testament to his productive and exceptional academic research skills. He is a high-calibre researcher, with 11 peer-reviewed publications. Mr. Leung led the HSV proof-of-concept project, and will be responsible for extending this work and conducting in vivo trials. He has excellent communication and leadership skills, making him an ideal candidate for this interdisciplinary project. Kevin Pennings, MASc Candidate. Mr. Pennings joined Dr. Filipe’s lab in 2013 during his undergraduate studies, and has since significantly developed his research skills and published 5 peer- reviewed papers. He is expected to defend his Master’s this August, and continue working on this project as a Research Assistant. Kevin has exclusive experience on pullulan, which makes him an asset for further developing this project. Marianne Chew, MSc. Ms. Marianne is a skilled lab technician at McMaster Immunology Research Center. She received various awards including the CIHR Frederick Banting and Charles Best Canada Graduate Scholarship. She will be hired temporarily (for 7 months), and will assist Mr. Leung with animal testing. 5 Filipe, C. I2I Proposal Section 2: Description of the Technology Transfer Plan MILO Contact Glen Crossley, Assistant Director, McMaster Industry Liaison Office (MILO) Phone: (905) 525-9140 x20372 Email: [email protected] 2.1 Embodiment of the Technology Work on this project will result in several prototypes that position our pullulan-based technology as an engineering solution platform for vaccine stabilization. Vaccine prototypes for influenza, BCG (tuberculosis) and tetanus will be stabilized and dosed in P/T films, and tested for performance in vitro and in vivo. This technology, in its final form, will essentially act as a carrier for reagents. The packaging of the product will be altered and presented in a tablet/capsule form, containing the necessary dose of ingredient(s). However, this technology integrates with existing user process and will not require new equipment nor extra or different steps. For example, the current process of preparing vaccines requires adding saline buffer to a vaccine powder before administering; in comparison, our proposed system requires the addition of saline buffer to a vaccine capsule/tablet instead. The end goal is to present these prototypes as strategic demonstration projects, to capitalize on interest shown by organizations specializing in vaccines, as evidenced by the letters of support attached. 2.2 Market Overview 2.2.1 Market Environment The market opportunity lies in the growing use of biological reagents for therapeutics, (“biologics”) and components for molecular diagnostics. One major market for this technology is the biopharmaceutical market, which deals with temperature and oxygen sensitive products. This industry is adding more such products to its repertoire, and depends on ‘cold’ chain logistics to handle storage and transportation needs. Unlike traditional “small-molecule” chemical drugs, biologic drugs require cold chain logistics require specialized cooling and transportation infrastructure, labour intensive management, and cost $100-$105 per square foot more than a conventional, ambient facility. Pullulan/trehalose (PT) stabilizing technology will address the needs of the target market by removing this costly dependency. This technology will act as the sole, cost-effective and user-friendly stabilizing method that will revolutionize how sensitive reagents are handled. Political Increasing regulations on temperature sensitive products are driving supply chain costs, according to the 8th UPS “Pain in the Chain” survey, which interviewed 421 healthcare logistics executives. The industry is in need for new innovation, and is responding through investments. According to the 2013 Cold Chain IQ survey, 68% of pharmaceutical and biotechnology manufacturers companies surveyed had organized budgets of over $10 million for innovation in temperature sensitive supply chain management. Economic The growing use of biological reagents such as biologic drugs is evidenced by growth in the biopharmaceutical market. The market for temperature sensitive drugs and biological products has a growth rate of 8% per year – double the growth rate of “conventional pharmaceuticals”. The drug market is estimated at $200 billion, with biotechnology-based products representing the majority of best selling pharmaceutical products by 2018. This signals a huge opportunity for our PT stabilizing technology to solve a growing need to reduce the costs of handling, storing and distributing biologics- based products. Movement towards health care equity will be met by eliminating the dependency of drug and vaccine delivery on a cold supply chain that does not reach the world’s remote and poor areas. Pullulan- 6 Filipe, C. I2I Proposal trehalose based technology will play a critical role in increasing market access to people who most need it. Technological Innovation in temperature and oxygen sensitive products (drugs and biologics) is driving the growth of these markets. The majority of R&D activities in human health are directed at both medical diagnostics and biopharmaceuticals, such as biotechnology-derived proteins, antibodies, enzymes, etc.. This trend presents a favorable demand forecast for stabilizing technologies. 2.2.2 Competitive Landscape and Barriers to Entry Competing methods for preserving biological reagents include lyophilization (freeze drying) and use of “proprietary formulations”. Lyophilization is commonly used to stabilize reagents, yet has many limitations. It is capital intensive, requires special equipment, and has high associated energy costs. Furthermore, dependency on a cold-supply chain is not eliminated, as many freeze-dried products still need to be refrigerated. The process also poses a risk on protein denaturation and membrane damage, and requires the addition of special “protective agents”. However, these additives can have adverse effects on overall product quality. Furthermore, products remain susceptible to damage from humidity and oxygen. Note that no major players dominate this market. Several emerging companies are exploiting research and utilizing proprietary formulations for stabilizing reagents. However, these companies operate in different markets. For example, US-based Biomatrica has a proprietary formulation based on the concept of anyhydrobiosis for preserving DNA, RNA and PCR-based assay reagents that can bypass cold-chain requirements. Customers are provided with the formulation and stabilize assay reagents in-house in an energy independent process. Canada-based DNA Genotek has patented processes for stabilizing DNA and RNA, with a focus on the sample collection market. Hence, there are no major players operating in this field that pose a threat to pullulan-based stabilizing technology operating in the drug/vaccine market. The major barrier to entry is heavy regulation on biopharmaceuticals, which will prolong time to market. One advantage of our systems is that both pullulan and trehalose are already FDA approved. In addition, market entry via collaboration projects (or licensing) with pharmaceutical companies should decrease barriers by capitalizing on their knowledge of compliance. Integration of technology into manufacturing assembly lines is also a barrier. However, cost of change is justified with the forecasted long-term cost-savings. The goal of demonstration projects with manufacturers is to provide a clearer picture of such cost-savings. 2.2.3 Competitive Advantage Vaccine stabilization based on pullulan-trehalose tackles the pain-points of the highlighted target market largely dependent on lyophilization and cold-chain logistics for storing, transporting, and handling vaccines. Pullulan is energy independent, does not require intensive capital investments, integrates with user processes and offers cost savings associated with handling substances at ambient conditions. Our proprietary knowledge includes casting and processability of pullulan, which allows packaging of pre-measured quantities of reagents. By focusing more on a “capsule/tablet” based format, the end-user can receive desired quantity of a vaccine from a licensed manufacturer, eliminating any need to handle vaccine in its sensitive form. Furthermore, the move into the biopharmaceutical market will unlock a plethora of challenges to solve, and will differentiate us from Biomatrica. This project will be used to further develop and position the technology as a platform designed as an engineering solution to vaccine stabilization. By showcasing the capability to integrate with an array of products, a technology license becomes much more valuable to interested companies. Note that this technology is well capable of integrating with DNA amplification methodologies such as loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA) and polymerase chain reaction (PCR). This will also make our technology relevant to the (growing) molecular diagnostic market. 7 Filipe, C. I2I Proposal 2.2.4 Overview of Costs The GMP pullulan mixture costs (excluding the cost of the vaccine itself) is approximately $23 for 1000 tablets. (Pullulan, the main ingredient, costs around $100 for 50 grams, which is enough for 12500 tablets). Thus, the stabilization mixture adds only 2.3 cents per vaccine. In regards to equipment, casting of pullulan requires an automated pipetting system with temperature control capabilities. As this pipetting system is assumed present in manufacturing facilities producing vaccines, it is not considered an additional cost to the production process. Furthermore, there are capital cost savings associated with eliminating the lyophilization process (freeze-drier), which is an additional cost saving advantage with our proposed system. As a major focus of this project will be scale-up, more accurate manufacturing costs will be provided moving forward. 2.3 Intellectual Property Strategy US (Serial No 15/034,914) and Canadian patent applications covering the creating of pullulan- based tablets for reagent preservation and assay simplification has already been filed. Our Freedom to Operate assessment, along with the international search report on the parent PCT application identified one US patent,which requires licensing technology from Auburn University. The patent from Auburn broadly claims preserving biological specimen at room temperature using pullulan only. We have had several converstations with their tech transfer office and they are please to work with us on licensing to third parties (if needed) or will simply continue to offer a non-exclusive “click through” license on their websited for $7500. A second patent has been filed by the Hayashibara Co. (Japan) on creating P/T films (EP1398346 A4), but the pullulan/trehalose ratios covered in that patent fall outside the ranges needed to preserve vaccines. Moreover, in that patent, the claims focus on the physical/mechanical properties of the films and not on the ability to preserve vaccines/viruses/phage. We are in the process of filing a new provisional patent to covering our specific formulations and believe we have the freedom to operate, and to further develop the commercial value of this technology. The McMaster Industry Liaison Office (MILO) is committed to commercialize this technology, and has been involved with licensing cooperation with Auburn University. 2.4 Commercialization Strategy The goal of this project is to further develop the commercial value of pullulan-based stabilizers. The commercial value of a pullulan-based stabilizer is dependent on its ability to integrate seamlessly with manufacturing processes, end-user processes, and an array of vaccines without altering performance or requiring many modifications. This will be done through focusing this project on user-validation (to refine how end-users use the product), processability (for scale-up), and prototyping. Prototypes will be developed with various vaccines to portray technological potential in the biopharmaceutical market. By positioning this technology as a platform capable of stabilizing several vaccines, a license becomes more valuable to biopharmaceutical manufacturers. At this moment, one pharmaceutical company (GSK) has expressed interest in this technology and provided a letter of support. The end goal is to turn their displayed interest into a technology license moving forward. We have also obtained a letter of support from Advanced Theranostics, which has an interest in using the technology to stabilize reagents for performing isothermal amplification of DNA. There is a clear market need (see Section 2.2.1) for innovation in vaccine stabilization, which aligns well with the forecasted growth in biopharmaceuticals. After the milestones of the project are met, the best commercialization pathway to reach market is through a technology license to pharmaceutical companies (specifically ones that develop vaccines). Given the nature of vaccines and the highly regulated environment, this is the fastest route to ensure our technology commercializes, meets a growing need, and plays a critical (social) role in increasing market access of vaccines. Our department will remain an innovative R&D hub, with plans to further expand this technology to encompass complex 8 Filipe, C. I2I Proposal biomolecules – another growing trend in the biopharmaceutical scene. McMaster University is committed to commercialize this technology, as displayed through prior investments for patenting our technology. Furthermore, McMaster University fully supports our approach, and has worked closely with us on developing agreements mentioned in the Intellectual Property Strategy section. 2.5 Benefit to Canada This project holds various socio-economic benefits to Canada. The pullulan-based stabilizing technology will benefit Canada’s well-established pharmaceutical market ($10.9 billion) through cost reduction in transportation, handling and storage of temperature-sensitive vaccines (and later other drugs). This will effectively increase the competitiveness of Canadian products. This technology will also increase market access globally as a result of liberation from cold chain requirements. Increasing market access and competitiveness of Canadian products is critical for sustaining manufacturing jobs in a highly globalized (and competitive) world. Furthermore, the maintenance of a cold supply chain accounts for 7% of the global hydrofluorocarbon consumption. Eliminating the use of a refrigerated supply chain through the PT stabilizing technology will curb these emissions. This has environmental benefits both on a local and global scale, and will reaffirm Canada’s role in tackling climate change. Furthermore, pullulan-based stabilizing technology reduces energy required during transportation of vaccines (and other biopharmaceutical) products. Research efforts for reducing national energy requirements align with Natural Resources Canada, where a significant part of their mandate is exploring ways to adapt and change distribution methods, in order to meet a growing energy demand sustainably. This technology has potential of revolutionizing energy-intensive supply chains, for the benefit of local, national and global populations. 9 Filipe, C. I2I Proposal References Ariel Schwarts. (2013, July) Fast Coexist. [Online]. http://www.fastcoexist.com/1682578/this-bill- gates-backed-super-thermos-saves-lives-with-cold-vaccines Dangerous Goods. (2012) Berlin Packaging. [Online]. http://www.berlinpackaging.com/system/resources/BAhbBlsHOgZmIlIyMDE0LzA5LzA1LzEzXz IwXzAwXzM4OV9TaGlwcGluZ19UZW1wZXJhdHVyZV9TZW5zaXRpdmVfTWF0ZXJpYWxz XzA4MTUyMDE0LnBkZg Jones Lang LaSalle. (2014) Perspectives on cold storage investment properties. Should you warm up to cold storage? [Online]. http://www.us.jll.com/united-states/en-us/Research/cold-storage- investment-perspectives-jll-nov2014.pdf?9c8bf36c-a6b5-4432-87e5-5149186c1c26 Michael J Akers, "Parenteral Preparations," in The Science and Practice of Pharmacy, 21st ed., David B Troy, Ed. Baltimore, United States of America: Lippincott Williams & Wilkins, 2006, pp. 802-829. Labconco. (2010) Cole Parmer. [Online]. http://www.coleparmer.ca/Assets/MoreInfo/Labconco_guide_freeze_dry_in_lab.pdf J-P Amorij, A Huckriede, J Wilschut, H W Frijlink, and W.L J Hinrichs, "Development of Stable Influenza Vaccine Powder Formulations: Challenges and Possibilities," Pharmaceutical Research, vol. 25, no. 6, pp. 1256-1273, June 2008. Ralf Otto, Alberto Santagostino, and Ulf Schrader. (2014, December) McKinsey & Company. [Online]. http://www.mckinsey.com/industries/pharmaceuticals-and-medical-products/our- insights/rapid-growth-in-biopharma Mark Lipowicz and Nicholas Basta. (2014, April) Pharmaceutical Commerce. [Online]. http://pharmaceuticalcommerce.com/supply-chain-logistics/2014-biopharma-cold-chain-forecast/ M. R. Rekha and Chandra P. Sharma, "Blood compatibility and in vitro transfection studies on cationically modified pullulan for liver cell targeted gene delivery," Biomaterials, vol. 30, no. 34, pp. 6655-6664, December 2009. United Parcel Service of America, Inc. (2015) United Parcel Service of America, Inc. [Online]. https://www.ups.com/media/en/UPS-PITC-Executive-Summary-North-America.pdf Cold Chain IQ. (2014) Global Cold Chain Management Solution: Insights Report 2014. Document. Bill & Melinda Gates Foundation. gatesfoundation.org. [Online]. http://www.gatesfoundation.org/What-We-Do/Global-Development/Vaccine-Delivery Wanis Kabbaj and Dirk Van Peteghem. (2015, February) PharmTech.com. [Online]. http://www.pharmtech.com/reinventing-cold-chain-high-stakes-market IBISWorld, "IBISWorld Industry Report: Global Biotechnology," IBIS World, 2016. Tse-Chao Hua, Bao-Lin Liu, and Haimei Zhang, Freeze-Drying of Pharmaceutical and Food Products. Cambridge, United Kingdom: Woodhead Publishing Limited, 2010. Markets and Markets, "Freeze Drying / Lyophilization Equipment Market by Scale (Lab, Pilot, & Industrial), Technology (Rotary, Manifold, & Tray - style) & by Accessories (Loading/Unloading System, Vacuum System, Clean-in-Place System, Control System) - Global Forecast to 2019 ," Markets and Markets, 2014. Biomatrica. (2016) Biomatrica. [Online]. http://biomatrica.com/company.php DNA Genotek. DNA Genotek. [Online]. http://www.dnagenotek.com/ROW/index.html Carlos Filipe, John D. Brennan, Robert Pelton, Sana Jahanshahi-Anbuhi, and Yingfu Li, "Method 10 Filipe, C. I2I Proposal of stabilizing molecules without refrigeration using water soluble polymers and applications thereof in performing chemical reactions ," Chemical WO2015066819 A1, May 14, 2015. Vitaly J. Vodyanoy et al., "Use of pullulan to isolate and preserve biological material," US7604807 B2, October 20, 2009. Toshiyuki Sugimoto and Yoshio Miyake, "Molded object having high pullulan content, process for producing the same, and use thereof ," Chemical EP1398346 A4, August 25, 2004. Sarah Turk, "IBISWorld Industry Report 32541bCA: Generic Pharmaceutical Manufacturing in Canada ," IBISWorld, 2016. Sarah Turk, "IBISWorld Industry Report 32541aCA: Brand Name Pharmaceutical Manufacturing in Canada ," IBISWorld, 2015. Jason Mathers. (2013, October) Environmental Defense Fund. [Online]. http://business.edf.org/blog/2013/10/16/cooling-the-climate-impact-of-the-cold-chain/ Natural Resources Canada. (2016, May) Natural Resources Canada. [Online]. http://www.nrcan.gc.ca/energy/energy-sources-distribution 11  9 (SNG, CRD, I2I, IRC, DND) Personal identification no. (PIN) Family name of applicant Valid 256630 Filipe INTELLECTUAL PROPERTY Complete this section if you need to discuss the plans for protecting and disposing of intellectual property arising from the grant. Do not exceed one page. McMaster University has filed the following patent application involving the use of pullulan-based mixture as a stabilizing agent. "Method for Stabilizing Various Molecules without Refrigeration Using Water Soluble Polymers - Demonstration Using Pullulan", US provisional patent application. 61/901,784, 2013. Filing Date: 2013-11-08 Any new IP arising from this project under I2I grant will be protected according to the normal policies of McMaster University and with cooperation from McMaster Industry Liaison Office. Form 101 (2011 W), page 9 of 9 PROTECTED WHEN COMPLETED Version française disponible  SEND ONE ORIGINAL ONLY DO NOT PHOTOCOPY APPENDIX C Referee Suggestions (Form 101) Complete Appendix C for all types of grants (except Discovery Grants, Research Tools and Instruments - Date Category 1, Major Resources Support Grants and Partnership Workshops Program). Read the instructions before completing the appendix. 2016/07/04 Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN) Filipe Carlos DM Valid 256630 Title of proposal Validation and Scale Up of Vaccine Stabilization for Ambient Handling Using a Pullulan/Trehalose Formulation Garnier, A (Alain) Area(s) of expertise A biochemical 1 Chemical Engineering Laval University engineering, Medicine: 2325 University Street Medical technology, Quebec, QC CANADA G1V0A6 Culture of cells for 1 (418) 6563106 therapeutic, Production PIN Lang. [email protected] of viral vaccines Haynes, C (Charles) Area(s) of expertise B Protein Purification, 2 Chemical and Biological Engineering The University of British Columbia Recombinant Proteins, 2360 East Mall Molecular Vancouver, BC CANADA V6T1Z3 Thermodynamics, 1 (604) 8225136 Chiral-Drug PIN Lang. [email protected] Purification, Biocompatible Area(s) of expertise C 3 PIN Lang. Area(s) of expertise D 4 PIN Lang. Area(s) of expertise E 5 PIN Lang. NSERC reviewing committee 1st committee reviewer Personal identification no. (PIN) 2nd committee reviewer Personal identification no. (PIN) 3rd committee reviewer Personal identification no. (PIN) Form 101, Appendix C (2011 W) PROTECTED WHEN COMPLETED Version française disponible