Production of monoclonal antibodies MCQs With Answer

Introduction: This quiz set on the production of monoclonal antibodies is designed for M.Pharm students to consolidate advanced knowledge in immunotechnology and biologics manufacturing. It covers core principles from hybridoma creation and recombinant expression to upstream process optimization, downstream purification, quality control and regulatory considerations. Questions emphasize mechanisms (fusion, selection, humanization), production platforms (CHO, NS0, transgenic), process modes (fed‑batch, perfusion), and critical quality attributes such as glycosylation, Fc‑mediated functions and aggregation. Practicing these MCQs will help students apply theoretical concepts to practical production challenges, improve problem‑solving for research and industry settings, and prepare for exams and professional roles in biopharmaceutical development.

Q1. What is the classical method used to generate monoclonal antibodies by fusing an antibody‑producing B cell with an immortal myeloma cell?

• Somatic hypermutation
• Hybridoma technology
• Phage display
• Transgenic animal expression

Correct Answer: Hybridoma technology

Q2. Which component of HAT medium is essential to select for successfully fused hybridoma cells by rescuing the salvage pathway?

• Hypoxanthine
• Aminopterin
• Thymidine
• Hemin

Correct Answer: Hypoxanthine

Q3. Why are myeloma partner cell lines used in hybridoma production typically HGPRT‑deficient?

• To enable secretion of higher affinity antibodies
• To confer resistance to aminopterin
• To allow selection in HAT medium only for fused cells
• To increase glycosylation complexity of antibodies

Correct Answer: To allow selection in HAT medium only for fused cells

Q4. Which fusion agent is most commonly used to promote cell fusion between spleen B cells and myeloma cells?

• Polyethylene glycol (PEG)
• Calcium phosphate
• Lipofectamine
• Electroporation buffer

Correct Answer: Polyethylene glycol (PEG)

Q5. Limiting dilution cloning is used during hybridoma development primarily to:

• Increase antibody glycosylation heterogeneity
• Obtain monoclonal populations from a single hybrid cell
• Transfect heavy and light chains into CHO cells
• Remove endotoxin from culture supernatant

Correct Answer: Obtain monoclonal populations from a single hybrid cell

Q6. Which analytical assay is most commonly used to screen hybridoma supernatants for antigen‑specific antibody production?

• Western blot for heavy chain only
• ELISA (enzyme‑linked immunosorbent assay)
• Flow cytometry for cell viability
• Mass spectrometry peptide mapping

Correct Answer: ELISA (enzyme‑linked immunosorbent assay)

Q7. In recombinant production of therapeutic mAbs, which cell line is the industry standard for large‑scale expression due to human‑like glycosylation and regulatory history?

• Escherichia coli
• CHO (Chinese Hamster Ovary) cells
• Saccharomyces cerevisiae
• Spodoptera frugiperda (Sf9) insect cells

Correct Answer: CHO (Chinese Hamster Ovary) cells

Q8. Which culture mode typically yields higher product concentration over a defined period and is widely used in commercial mAb manufacturing?

• Continuous perfusion without cell retention
• Fed‑batch culture
• Batch culture with daily harvests
• Solid‑phase fermentation

Correct Answer: Fed‑batch culture

Q9. What is the principal affinity ligand used in the primary capture step for IgG monoclonal antibodies during downstream purification?

• Protein G
• Protein A
• Protein L
• Concanavalin A

Correct Answer: Protein A

Q10. Which post‑translational modification on the Fc region most directly affects antibody-dependent cellular cytotoxicity (ADCC)?

• Deamidation of asparagine in variable region
• Core fucosylation of N‑linked glycans
• Oxidation of methionine in hinge
• C‑terminal lysine clipping

Correct Answer: Core fucosylation of N‑linked glycans

Q11. Humanization of a murine monoclonal antibody by CDR grafting primarily aims to:

• Increase antigen affinity by introducing somatic mutations
• Reduce immunogenicity while preserving antigen specificity
• Eliminate all N‑linked glycosylation sites
• Enable expression in bacterial systems

Correct Answer: Reduce immunogenicity while preserving antigen specificity

Q12. Which of the following methods generates fully human monoclonal antibodies without mouse sequences by using human antibody gene libraries displayed on bacteriophages?

• Transgenic mouse immunization
• Phage display
• Hybridoma from human B cells only
• Somatic hybridization with human myeloma

Correct Answer: Phage display

Q13. For aseptic formulation of mAbs, removal of host cell proteins (HCP) is critical because HCPs can:

• Enhance FcRn binding and increase half‑life
• Serve as active pharmaceutical ingredients
• Induce immunogenicity and affect stability
• Improve antibody aggregation profile

Correct Answer: Induce immunogenicity and affect stability

Q14. Which viral clearance step is commonly integrated into downstream processing to ensure removal/inactivation of adventitious viruses?

• Tangential flow filtration only
• Low‑pH viral inactivation and nanofiltration
• Protein A affinity chromatography exclusively
• Ion exchange chromatography without depth filters

Correct Answer: Low‑pH viral inactivation and nanofiltration

Q15. FcRn binding enhancement is engineered into some therapeutic antibodies to:

• Increase complement activation
• Prolong serum half‑life through neonatal Fc receptor recycling
• Improve tissue penetration by reducing size
• Increase glycosylation heterogeneity

Correct Answer: Prolong serum half‑life through neonatal Fc receptor recycling

Q16. Which quality attribute is assessed by size‑exclusion chromatography during release testing of mAbs?

• Charge heterogeneity
• Aggregate content and monomer purity
• Host cell DNA levels
• N‑linked glycan identity

Correct Answer: Aggregate content and monomer purity

Q17. In choosing an expression system for a mAb with complex human‑like glycosylation, which system is least appropriate?

• CHO cells
• NS0 murine myeloma cells
• Escherichia coli
• HEK293 human embryonic kidney cells

Correct Answer: Escherichia coli

Q18. Which regulatory concept is essential when developing a biosimilar monoclonal antibody to demonstrate similarity to the reference product?

• Identical cell bank and manufacturing site as originator
• Extensive analytical comparability and clinical justification
• Use of the same mouse immunization protocol
• No requirement for pharmacokinetic studies

Correct Answer: Extensive analytical comparability and clinical justification

Q19. What is the main purpose of using serum‑free, chemically defined media in mAb production?

• To increase endotoxin levels for potency
• To reduce variability and risk of adventitious agents
• To guarantee faster microbial growth
• To increase glycan heterogeneity for better efficacy

Correct Answer: To reduce variability and risk of adventitious agents

Q20. Glycoengineering to remove core fucose from Fc glycans is often performed to:

• Reduce antibody binding to antigen
• Enhance ADCC activity by improving FcγRIIIa binding
• Prevent FcRn recycling and shorten half‑life
• Increase complement activation via C1q binding only

Correct Answer: Enhance ADCC activity by improving FcγRIIIa binding

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