Biologics Production and Contamination Quiz

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Questions and Answers

What is the primary source of contamination for human cells during production?

Environmental factors

Sero-derived components

The operating personnel (correct)

The cell line itself

Which method is most commonly used to detect contamination in biological products?

Culture methods

Immunofluorescence assays

Polymerase chain reaction (PCR) (correct)

Nucleic acid sequencing

What can happen if a contaminant is detected during the manufacturing process?

The manufacturing is suspended (correct)

The production continues as normal

Products are released to the market with caution

Immediate actions to quarantine the serum

What is a key reason for not using animal serum in the production of biologics?

It is a common source of contamination (C) Signup and view all the answers

Which viral assay is mentioned as a method for detecting viral contamination?

In vitro viral assay (IVV) (C) Signup and view all the answers

What usually mitigates the risks associated with viral contamination in biologics?

Quality control protocols (A) Signup and view all the answers

In which process levels can contamination events occur?

Both upstream and downstream, and other unidentified stages (A) Signup and view all the answers

What is the implication of using only human cell lines in biologics production?

It poses a higher contamination risk (A) Signup and view all the answers

What is the starting point of the glycosylation pathway in a CHO cell line?

Endoplasmic reticulum (A) Signup and view all the answers

Which component is crucial for the quality and stability of therapeutic glycoproteins?

Sialic acid (A) Signup and view all the answers

How does the presence of galactose influence glycoproteins?

It is important for their plasma half-life. (B) Signup and view all the answers

What can influence the activity of the enzymes involved in glycosylation?

Cell culture parameters (B) Signup and view all the answers

What role do receptors in the liver play concerning glycoproteins?

They bind to non-sialylated glycoproteins for removal. (D) Signup and view all the answers

The final form of a glycoprotein is primarily determined by which of the following?

Glycosylation levels and terminal sugars (B) Signup and view all the answers

Which of the following factors could NOT be a parameter to control the glycosylation pathway?

Temperature of the cytoplasm (C) Signup and view all the answers

What is a characteristic of the enzymes involved in the glycosylation pathway?

Their activity depends on the environmental conditions. (D) Signup and view all the answers

What can lead to toxicity in pharmaceutical formulations?

Materials in contact with the protein (A) Signup and view all the answers

Which factor is NOT mentioned as a cause of protein contamination?

Chemical stability of containers (D) Signup and view all the answers

What was identified as a potential cause for immunogenic responses in preparations of epoietin?

Phenolic derivatives from rubber stoppers (C) Signup and view all the answers

Why must the conditions during protein handling be closely monitored?

To prevent unwanted modifications to the protein (B) Signup and view all the answers

What does the term 'heterogeneity' refer to in the context of protein therapeutics?

Presence of different protein isoforms (D) Signup and view all the answers

What happens if the properties of protein variants differ significantly from the desired product?

They are classified as impurities (A) Signup and view all the answers

What does the term 'process related impurities' refer to?

Impurities introduced during the manufacturing process (A) Signup and view all the answers

What aspects can influence the activity and toxicity of biopharmaceuticals?

Storage conditions and vial materials (C) Signup and view all the answers

What is the primary purpose of Process Analytical Technology (PAT)?

To ensure timely measurements in manufacturing processes (A) Signup and view all the answers

What is the European Pharmacopoeia's role in relation to protein therapeutics?

To outline guidelines for acceptable purity levels (B) Signup and view all the answers

Which of the following accurately describes at-line measurement?

Measurements performed on samples removed from the production line (D) Signup and view all the answers

Which organization provides guidelines for product quality control beyond the pharmacopeia?

EMA (B) Signup and view all the answers

What is indicated by the title of a protein in terms of yield?

Concentration of protein per unit volume (C) Signup and view all the answers

What distinguishes online measurement from inline measurement?

Inline measurement occurs without interruption in flow (C) Signup and view all the answers

What is a common consequence of the sample returning to its original container during measurement?

Contamination of the sample (D) Signup and view all the answers

What does the EMA aim to ensure through its guidelines?

Maintenance of product quality and safety (A) Signup and view all the answers

Why is serum banned in mammalian cell culture when producing proteins for patient injection?

It may contain viruses that can be harmful. (C) Signup and view all the answers

What is the role of DHFR in the context of gene expression and cell growth?

It blocks the production of folic acid. (D) Signup and view all the answers

What advantage do CHO-S cells provide in culture conditions?

They can grow in suspension culture without serum. (A) Signup and view all the answers

How does methotrexate function in relation to DHFR and cancer treatment?

It inhibits DHFR, preventing cell replication. (A) Signup and view all the answers

In the context of selecting clones for protein production, what is the purpose of measuring DHFR expression?

To establish a system that correlates with protein expression. (C) Signup and view all the answers

What is a primary function of stirred tank bioreactors in cell culture?

To mix and agitate the culture medium (C) Signup and view all the answers

What is one consequence of using serum in cell culture that affects research reproducibility?

Serum components vary significantly between batches. (D) Signup and view all the answers

Which of the following best describes the importance of sterilization in bioreactors?

To ensure the quality of the final protein product (C) Signup and view all the answers

What is the primary goal of coupling the gene for the desired protein with the DHFR gene?

To allow for gene amplification based on DHFR expression. (A) Signup and view all the answers

What happens to cells that express higher levels of DHFR when exposed to methotrexate?

They grow better than other clones. (B) Signup and view all the answers

What does process characterization aim to achieve?

To develop a process for routine commercial manufacturing (A) Signup and view all the answers

Which step typically follows cell harvesting in the bioprocessing of biologics?

Downstream processing (A) Signup and view all the answers

What type of chromatography is considered typical for the downstream processing of mammalian cells?

Capture chromatography (C) Signup and view all the answers

What is the role of process verification in manufacturing?

To confirm the effectiveness of the final manufacturing process (A) Signup and view all the answers

Why is virus removal particularly critical in the bioprocessing of mammalian cells?

Mammalian cells are often already infected or susceptible to infection (C) Signup and view all the answers

Which of the following is a component of the classic bioprocessing scheme?

Cell harvesting (A) Signup and view all the answers

Study Notes

Protein Denaturation and Renaturation

Denaturation can be good, as it allows misfolded proteins in bacteria to be further denatured and then renatured.
Glycosylation
Virus
Pirogens can be contaminants in eukaryotic cells.
Recombinant proteins in E. coli.
E. coli is a common host for producing heterologous proteins.
Glycosylation is a fundamental post-translational process
Bacteria can't modify proteins with glycans, eukaryotes can.
Different glycans attached to a protein creates variation
DNA is messenger RNA!

Protein Expression in E. coli

Many reviews exist on recombinant protein production in bacteria
Protein production is important for drug discovery and development.
Genetically engineered yeast strains allow for mammalian-like glycosylation patterns.
CHO and NSO cells are commonly used for mAb production.

Glycosylation

Glycosylation is a fundamental post-translational process.
Bacteria can't perform glycosylation.
Eukaryotes can modify proteins with glycans, leading to diverse protein variations.

Sialic Acid

Sialic acid
Important for the serum half-life of glycoproteins
Prevents degradation and removal from the blood.
Prevents antigenic determinants from exposing glycoproteins.
Influencing the thermal stability, its resistance to proteolytic degradation and its solubility.

Industrial Bioprocessing of Biologics

Cell harvesting and culture
Microbial purity is important for prokaryotic systems.
Virus-free systems are important for mammalian cell cultures, which often utilize viral removal steps.
Typical cell culture production schemes range from small-scale T-flasks to large-scale bioreactors.
Fed-batch fermentation is a method to extend fermentation time due to added nutrients.
Continuous fermentation allows for continuous product extraction, but introduces risk of contamination.

Culture Media and Other Materials

Media quality and traceability are important in bioprocessing.
Contaminants in the media can arise from formulation or substances used.
Extraction and purification of resulting products depend on used cells and proteins.

Process Characterization

The process must be designed to be suitable for routine commercial manufacturing.
Process development is the stage to reach a potential process design.
Process evaluation involves performing steps on a small and/or commercial scale.
Process verification studies are to confirm manufacturing process effectiveness.

Downstream Processing

Four main sections: primary recovery, viral clearance, purification, and formulation.
Primary recovery involves removing contaminating components.
Viral Clearance is to remove viruses from mammalian cell-derived proteins.
Purification ensures that the product meets purity specifications.
Formulation is to mix the product with appropriate excipients.

Important Concepts in Bioprocessing

Mammalian cell cultures are often grown in stirred tank bioreactors.
Sterilization methods (physical and chemical) are crucial to eliminating contaminants.
Process characterization involves designing processes suitable for commercial manufacturing.
Process validation confirms efficiency and consistency.

Cell Culture Process Design

Robust mammalian cell lines with high expression capabilities and large-scale continuous stirred tank reactor (CSTR) formats are common and preferred.
High throughput and process-oriented solutions (scale-up, cleaning, sterilization) are required.
Cell culture process design, robust and reproducible mammalian cell culture processes, suspension, and/or batch processes.
Gene-amplification and host cell protein removal steps are important.

Cell Line Transfection and Selection

CHO cells are the most utilized mammalian host cells.
CHO, NSO, BHK, HEK-293, and PER-C6 are alternative cell lines.
These cells are adapted for suspension culture.
Virus removal is common in these systems.

Additional Notes on the topic

The use of DHFR strains and MTX induced gene amplification are important aspects for amplification of the target gene.
Upstream processes can be optimized using high expression vector systems, cell lines capable of high product output per day, optimized media, quality of instrumentation and process conditions.
Crucial factors for cell culture optimization include, but are not limited to, cell and physical parameters, biochemical parameters, processes and technology.

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Description

Test your knowledge on the key aspects of contamination in biologics production. This quiz covers various sources of contamination, detection methods, and the implications of using human cell lines. Assess your understanding of best practices in the field of biomanufacturing.