Biopharmaceuticals and Cell Growth Media

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Questions and Answers

What are primary metabolites, and why are they essential during the growth phase of cells?

Primary metabolites are essential compounds such as amino acids and vitamins synthesized by cells during their growth phase, crucial for their growth and development.

List two examples each of sugars and vitamins that are commonly used in growth media for mammalian cells.

Examples of sugars include glucose and lactose; examples of vitamins include ascorbic acid and thiamine.

Describe the lag phase of bacterial growth during fermentation.

The lag phase is when bacteria do not divide but adapt to the growth conditions in the medium, preparing for subsequent growth.

What occurs during the stationary phase of bacterial growth?

During the stationary phase, active cell growth slows due to nutrient depletion and medium spoilage, leading to stable cell numbers. Signup and view all the answers

Identify the trace minerals necessary for mammalian cell growth, and explain their role.

Essential trace minerals include iron, copper, cobalt, and magnesium, which play vital roles in enzyme function and cellular metabolism. Signup and view all the answers

What is the primary source for the production of biopharmaceutical products as of 2010?

Mammalian cells. Signup and view all the answers

Identify the three classifications of bioreactor systems based on capacity.

Laboratory (< 50 L), Experimental (50 – 50,000 L), Industrial (> 50,000 L). Signup and view all the answers

How long do cultured cells remain in bioreactors during continuous processes?

90 days. Signup and view all the answers

What happens to the reactor after the biochemical conversions in batch processes?

The tank is emptied and the end products are purified. Signup and view all the answers

What is the purpose of the starting material and cells in bioreactors?

They are used for biochemical conversions to produce therapeutic recombinant proteins. Signup and view all the answers

What percentage of the 58 biopharmaceutical products were produced in E. coli from 2006 to 2010?

Approximately 29% (17 out of 58). Signup and view all the answers

Which type of bioreactor system is used for small-scale experimentation?

Laboratory bioreactor. Signup and view all the answers

In large-scale manufacturing, what is the role of fermenters?

Fermenters facilitate the cultivation of cells for producing therapeutic recombinant proteins. Signup and view all the answers

What type of bioreactor system is defined by a capacity greater than 50,000 L?

Industrial bioreactor. Signup and view all the answers

What is the significance of having fermentation or bioreactor processes in biotechnology?

They are essential for the large-scale production of biologics and therapeutic proteins. Signup and view all the answers

What is biotechnology and how does it utilize living organisms for production processes?

Biotechnology is a multidisciplinary field that integrates natural and engineering sciences to apply organisms for product and service creation through processes involving living organisms like bacteria and yeast. Signup and view all the answers

Explain the role of genetic engineering in modifying organisms and provide an example.

Genetic engineering involves altering the genetic makeup of organisms by inserting genes from one into another to achieve desired traits; for instance, creating bacteria that produce human insulin. Signup and view all the answers

What are recombinant proteins and why are they significant in biotechnology?

Recombinant proteins are artificially produced using genetic engineering techniques, and they are significant in biotechnology for their use in research and as drugs for treating diseases. Signup and view all the answers

Describe the process through which recombinant proteins are produced using a common bacterium.

The process involves inserting a human gene into the genetic material of a bacterium, which then utilizes this genetic information to produce the encoded protein. Signup and view all the answers

When was the first recombinant protein used in treatment and what was it?

The first recombinant protein used in treatment was recombinant human insulin, introduced in 1982. Signup and view all the answers

What impact has the recombinant protein industry had since its inception?

The recombinant protein industry has rapidly grown, with over 130 recombinant proteins approved by the US FDA for clinical use to date. Signup and view all the answers

Identify two applications of biotechnology in the production of valuable substances.

Biotechnology is used to produce vaccines and enzymes through the use of engineered organisms. Signup and view all the answers

What are some drawbacks of using animal sources for protein pharmaceuticals?

Variability in amino acid sequences, limited production quantities, low purity, and potential immunological reactions. Signup and view all the answers

List the first two steps involved in recombinant protein production.

Isolating the gene and vector, and combining the gene with the vector. Signup and view all the answers

What is the significance of integrating the recombinant vector into the host cell?

It allows for the expression of the desired protein within the host organism. Signup and view all the answers

Identify one potential contaminant that may need to be removed in downstream processing.

Endotoxins or residual host cell proteins. Signup and view all the answers

Why is the choice of expression systems important in biotechnology?

Different expression systems can affect yield, post-translational modifications, and overall protein functionality. Signup and view all the answers

What type of cultivation system is typically used for large-scale recombinant protein production?

Bioreactors. Signup and view all the answers

What role does the culture medium play in recombinant protein production?

It provides essential nutrients for the host cells to grow and produce proteins. Signup and view all the answers

What is downstream processing in biotechnology?

The purification and recovery of biological products after fermentation or cell culture. Signup and view all the answers

What is one of the major benefits of recombinant protein production compared to traditional methods?

Recombinant production allows for larger quantities of proteins with high purity levels. Signup and view all the answers

What is the doubling time of E. coli under optimal conditions?

About 20 minutes. Signup and view all the answers

List one significant advantage of using mammalian cells in biotechnology.

They provide post-translational modifications like glycosylation. Signup and view all the answers

Name one limitation of using mammalian expression systems in industrial applications.

They are highly expensive. Signup and view all the answers

What type of host organism is E. coli classified as?

Prokaryotic cells. Signup and view all the answers

What does 'scalability' refer to in the context of mammalian cell use in biotechnology?

The ability to produce proteins in both small or large-scale processes. Signup and view all the answers

Identify the most commonly used host cell line for therapeutic protein production.

Chinese Hamster Ovary (CHO) cells. Signup and view all the answers

Explain one reason why rich complex media are advantageous for E. coli cultivation.

They can be made from inexpensive and readily available components. Signup and view all the answers

What is the main advantage of plasmid transformation in E. coli?

It can be performed in as little as 5 minutes. Signup and view all the answers

Why might potential contamination with animal viruses be a concern in mammalian expression systems?

It can affect the safety and efficacy of therapeutic products. Signup and view all the answers

Mention one reason why proteins produced in mammalian cells are preferred in human applications.

They are more likely to be bioactive and functional in humans. Signup and view all the answers

Study Notes

Course Information

Course title: Biochemistry and Biotechnology Fundamentals, Industrial Processes in Biotechnology
Course code: 1120-111
Instructor: Dr. Ahmed Hemdan
Department: Pharmaceutical Chemistry

Biotechnology

Multidisciplinary field integrating natural sciences and engineering
Organisms used to create products and services (genetic engineering)
Involves producing valuable substances from raw materials using living organisms (bacteria, yeast)

Recombinant Proteins

Artificially produced using genetic engineering
Key role in biomedical biotechnology
Used in research and as drugs for various diseases
Produced by inserting a human gene into a bacterium
Recombinant human insulin was the first such protein used in treatment in 1982
Over 130 recombinant proteins are currently FDA-approved for clinical use
Recombinant proteins used clinically include hormones, interferons, interleukins, growth factors, etc., for treating diabetes, dwarfism, and other diseases.

Plasmids

Small, circular DNA molecules
Found in bacteria and other microorganisms
Physically distinct from chromosomal DNA
Replicate independently

Genes

Specific DNA sequences
Contain information to code for functional products (usually proteins or RNA molecules)
Human protein-coding genes vary widely in length from a few hundred to millions of bases
Shortest genes are tRNA genes (70-90 nucleotides)
Human insulin gene (INS) is a relatively small gene with 1,425 base pairs.

Protein Pharmaceuticals from Animal Sources

Previously, protein pharmaceuticals/hormones were derived from animals (e.g., bovine or porcine)
Potential drawbacks include: not having the exact same amino acid sequence as in humans; difficulty obtaining large quantities; limited purity, and immunological reactions.

Recombinant Protein Production Steps

1. Isolate the gene and vector (e.g., bacterial plasmid)
2. Combine the gene and vector
3. Integrate the recombinant vector into host cells
4. Grow the cells in a culture medium
5. Extract the protein

Large-Scale Production of Biotechnology Products (Pharmaceuticals)

Expression Systems: (choosing the appropriate organism eg. Bacteria, Yeast, Filamentous fungi, Mammalian cells, and Unicellular algae).
Cultivation Systems: Large tanks also known as fermenters or bioreactors.
- Includes Laboratory-scale systems (<50L)
- Experimental systems (50-50,000L)
- Industrial systems (>50,000L)
Medium of cultivation: The liquid mixture of nutrients and salts for cells to grow.
Removal of Potential Contaminants: Crucial for product purity
Downstream Processing: Purification and recovery of biological products.

Three Bioreactor Processes

Discontinuous/Batch Process: Material added to tank and processes till completion, then tank emptied and product purified
Continuous Process: Continuous supply of raw materials, while equivalent amount of the reaction mixture containing the end product is taken out
Semi-continuous Process: Cells remain in the reactor for a fixed period and are supplied with fresh medium and starting material regularly

Cultivation Systems (Mammalian Cells) - Components of Growth Medium

Sugars: Glucose, lactose, sucrose, and maltose
Fats: Fatty acids and triglycerides
Water: Sterile water (high quality)
Amino acids: Glutamine
Electrolytes: Calcium, sodium, potassium, and phosphate
Vitamins: Ascorbic acid, tocopherol, thiamine, riboflavin, and folic acid
Fetal Calf Serum: proteins, albumin, transferrin
Trace Minerals: Iron, copper, cobalt, and magnesium
Hormones: Growth factors

Cultivation Systems (Bacteria) - Phases of Growth

Lag Phase: Cells adapt to the medium; no cell division
Log Phase: Rapid cell growth and gene expression
Stationary Phase: Cell growth slows; cell division and death rates are approximately equal.
Death Phase/Decline Phase: Nutrient depletion and waste accumulation leads to cell death.

Primary Metabolites

Synthesized during the growth phase
Essential for growth (e.g., amino acids, vitamins)
Amount increases as long as cells are proliferating

Secondary Metabolites

Produced at late stages of cell life cycle
Important for various applications, including antibiotics and toxins (but need conditions to be optimal)

Optimization of Recombinant Protein Production

Reduce lag phase duration
Delay stationary phase onset
Control culture medium pH
Effective control of oxygen and temperature
Strict sterile measures and work protocols

Potential Contaminants in Cultivation Process

Viral contaminants: Removed by chromatography, inactivation (heat or neutralization).
Bacterial contaminants: Removed by chromatography.
Cellular DNA contaminants: Removed.
Foreign protein contaminants: These pose health risks through immune reactions.

Therapeutic Proteins produced by Recombinant Protein Technology-Criteria

Ease of large-scale production and purification.
Absence of peptides and other contaminants.
Potentially fewer immune reactions.
Meeting regulatory standards.

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Description

This quiz explores essential concepts related to biopharmaceutical production, including primary metabolites and their role in cell growth. It also delves into the phases of bacterial growth during fermentation and the importance of trace minerals in mammalian cell culture. Test your understanding of bioreactor systems and the biopharmaceutical products derived from E. coli.