Identifying a Potential Biotechnology Product PDF

Identifying a Potential Biotechnology Product Chapter 6  Learning Outcomes ! Give examples of biotechnology products derived from plant and animal sources and discuss the challenges of extracting compounds ! Identify the steps in a Comprehensive Product Development Plan and use it to...

Identifying a Potential Biotechnology Product Chapter 6  Learning Outcomes ! Give examples of biotechnology products derived from plant and animal sources and discuss the challenges of extracting compounds ! Identify the steps in a Comprehensive Product Development Plan and use it to determine whether a potential biotechnology product is worth manufacturing ! Discuss the types of assays done as potential products move through process development and identify the additional assays required for pharmaceutical development ! Describe how an ELISA or a Western blot is conducted and what the results of each assay can reveal ! Explain how scientists test the effectiveness of antibiotics and antimicrobials and discuss the significance of antibiotic resistance. ! Describe the role of CHO cells in protein product development ! Describe the typical recombinant DNA protein product pipeline, additional steps required by the FDA for pharmaceutical proteins, and possible formulations of the final product 6.1 Sources of Potential Products For thousands of years, people have used plant and animal organs for many purposes. Recently, scientists have learned how to use plant and animal parts as sources of products: Pancreas of livestock is used to make insulin. Foxglove plant is used to regulate irregular heartbeat. Harnessing the Potential of Materials Produced in Nature Sometimes a product is made in very small quantities in nature so it must be produced Modeling the Research and Development of a Potential Product Estimating market size Identifying product sources Creating a Comprehensive Product Development Plan (CPDP) 1. Does the product meet a critical need? Who will use the product? 2. Is the market large enough to produce sufficient sales? How many customers are there? 3. Does preliminary data support that the product will work? Will it do what the company claims? 4. Can patent protection be secured? Can the company prevent other companies from producing it? 5. Can the company make a profit on the product? How much will it cost to make it? How much can it be sold for? 6.2 The Use of Assays If a substance is chosen as a potential product, researchers must be able to test for its presence, activity, and concentration. The product must be “assayed.” Other assays are done for potency, toxicity, and stability. Assays are performed at every step in the development of a product. The 24-well plate reveals samples with different amounts of amylase activity. Amylase breaks down starch to sugar. In the activity assay, as amylase breaks down starch, the dark color of a starch/iodine mixture becomes lighter. The lighter the color, the more active the amylase. Decreasing amounts of protein are indicated by less blue color in the tube. The right-hand tube is a negative control that contains no protein. The tubes to the left have increasing concentrations of protein.  6.3 Enzyme Linked Immunosorbent Assay (ELISA) One of the most important and frequently used molecular assays is the ELISA. An ELISA is very specific and will recognize a single type of protein or other antigenic molecule in a mixture of others. ELISA utilizes two important phenomenon: 1) antibody antigen specificity to recognize only very specific molecules 2) enzyme activity on colorimetric reagents for visualization purposes. In an ELISA, the goal is to recognize the antigen and measure its concentration.  6.4 Western Blots In a Western blot, samples are run on a PAGE gel and then the protein bands are transferred to a blotting membrane (PVDF or nitrocellulose). The visualization of a Western blot can confirm the presence of a particular protein at very low concentrations. Western Blot Gel/Transfer Membrane Setup Diagram. During a Western blot, electrical current carries protein bands from the PAGE gel to the blot transfer membrane. 6.5 Looking for New Products in Nature Herbal remedies from the rainforest Nutraceuticals is a term used for herbal remedies that come from food and plants that may have pharmaceutical or nutritional benefits. Recently there has been interest in the possible antimicrobial activity of many plants extracts. Herbal dietary supplements are controversial because many are not approved by the FDA and evidence of effectiveness is anecdotal. 6.6 Producing Recombinant DNA (rDNA) Protein Products rDNA was first produced in the mid 1970s. Recombinant refers to the process of combining DNA from two sources.