Biologics: recombinant hormones/proteins MCQs With Answer

Introduction: Biologics: recombinant hormones/proteins MCQs With Answer is a concise study resource tailored for M.Pharm students studying Biological Evaluation of Drug Therapy. This quiz set covers core concepts including expression systems, post‑translational modifications, formulation and stabilization strategies, analytical characterization, immunogenicity, potency assays, and regulatory aspects specific to recombinant hormones and therapeutic proteins. Questions are designed to test applied understanding and decision‑making relevant to laboratory practice, bioprocessing and clinical evaluation. Use these MCQs to reinforce theory, prepare for exams, and identify areas for deeper study in biologics development, quality control and therapeutic evaluation.

Q1. Which expression system has historically been the primary host for large‑scale production of recombinant human insulin?

• Escherichia coli
• Saccharomyces cerevisiae
• CHO (Chinese Hamster Ovary) cells
• Insect cell (baculovirus) system

Correct Answer: Escherichia coli

Q2. Why is glycosylation critical for the in vivo activity of recombinant erythropoietin (EPO)?

• It creates disulfide bonds required for receptor binding
• It increases solubility but does not affect half‑life
• It is essential for proper folding in the cytosol
• It prolongs circulation half‑life and affects receptor interaction and stability

Correct Answer: It prolongs circulation half‑life and affects receptor interaction and stability

Q3. Which type of assay is most appropriate to determine the biological potency of a recombinant growth factor?

• Competitive ligand binding ELISA
• Cell‑based bioassay measuring proliferation or signaling
• SDS‑PAGE followed by Coomassie staining
• Endotoxin Limulus Amebocyte Lysate (LAL) test

Correct Answer: Cell‑based bioassay measuring proliferation or signaling

Q4. What is the primary pharmacokinetic benefit of PEGylating a therapeutic protein?

• Increases enzymatic activity by altering the active site
• Promotes glycosylation to improve receptor binding
• Reduces renal clearance and increases plasma half‑life
• Targets the protein to lysosomes for faster clearance

Correct Answer: Reduces renal clearance and increases plasma half‑life

Q5. Which analytical method is commonly used to monitor residual host cell proteins (HCPs) in recombinant protein preparations?

• UHPLC‑MS for total protein mass
• Western blot for target protein only
• Host‑cell protein specific ELISA
• Size‑exclusion chromatography with UV detection

Correct Answer: Host‑cell protein specific ELISA

Q6. What contaminant is most commonly responsible for pyrogenic reactions associated with recombinant protein products?

• Residual host DNA
• Endotoxin (lipopolysaccharide) from Gram‑negative bacteria
• Cell culture serum albumin
• Non‑ionic surfactant excipients

Correct Answer: Endotoxin (lipopolysaccharide) from Gram‑negative bacteria

Q7. Fc‑fusion proteins have extended half‑life primarily because the Fc domain engages which mechanism?

• Enhances renal filtration through albumin binding
• Increases proteolytic cleavage in blood
• Recycles via the neonatal Fc receptor (FcRn) preventing lysosomal degradation
• Directs proteins to adipose tissue depots for slow release

Correct Answer: Recycles via the neonatal Fc receptor (FcRn) preventing lysosomal degradation

Q8. Which of the following is a major risk associated with aggregation of therapeutic proteins?

• Improved receptor specificity
• Reduced viscosity at high concentration
• Increased immunogenicity and potential neutralizing antibody formation
• Enhanced glycosylation heterogeneity

Correct Answer: Increased immunogenicity and potential neutralizing antibody formation

Q9. Which chromatographic technique is best suited to separate protein species based on hydrodynamic radius (size)?

• Anion‑exchange chromatography (AEX)
• Hydrophobic interaction chromatography (HIC)
• Size‑exclusion chromatography (SEC)
• Affinity chromatography using immobilized metal ions

Correct Answer: Size‑exclusion chromatography (SEC)

Q10. For recombinant glycoprotein therapeutics, which critical quality attribute must be characterized because it influences biological activity and clearance?

• Amino acid sequence only, glycosylation is irrelevant
• Glycosylation profile (site occupancy and glycoforms)
• Bulk pH of the final container only
• Concentration of non‑ionic surfactant exclusively

Correct Answer: Glycosylation profile (site occupancy and glycoforms)

Q11. What is a common limitation when expressing human glycoproteins in Escherichia coli?

• Excessive glycosylation compared with human proteins
• Inability to perform eukaryotic N‑linked glycosylation
• Overproduction of chaperones that hyperfold proteins
• Excessive formation of disulfide bonds in the cytoplasm

Correct Answer: Inability to perform eukaryotic N‑linked glycosylation

Q12. Which approach is most appropriate to detect neutralizing anti‑drug antibodies that block biological activity of a therapeutic protein?

• Direct sandwich ELISA measuring binding antibodies only
• Western blot for protein size changes
• Cell‑based neutralization assay that measures functional inhibition
• Mass spectrometry quantification of serum drug concentration

Correct Answer: Cell‑based neutralization assay that measures functional inhibition

Q13. Regulatory approval of a biosimilar typically requires which demonstration compared with the reference product?

• Only identical amino acid sequence, without analytical comparability
• Comprehensive comparability including analytical, nonclinical and clinical studies showing no clinically meaningful differences
• Complete new clinical development program identical to an originator biologic
• No requirement for immunogenicity assessment

Correct Answer: Comprehensive comparability including analytical, nonclinical and clinical studies showing no clinically meaningful differences

Q14. What is the primary goal of codon optimization when designing a gene for expression in a heterologous host?

• Introduce rare codons to slow translation and improve folding
• Maximize use of host‑preferred codons to increase translation efficiency
• Ensure the protein is glycosylated in E. coli
• Change amino acid sequence to increase immunogenicity

Correct Answer: Maximize use of host‑preferred codons to increase translation efficiency

Q15. For many refrigerated recombinant protein therapeutics, what is the recommended storage temperature range to maintain stability?

• Room temperature (20–25 °C)
• Frozen at −80 °C only
• Refrigerated at 2–8 °C
• Warm storage at 37 °C to simulate body temperature

Correct Answer: Refrigerated at 2–8 °C

Q16. Disulfide bonds in therapeutic proteins primarily contribute to which property?

• Increasing the net negative charge only
• Stabilizing tertiary and quaternary structure through covalent crosslinks
• Enhancing glycosylation site occupancy
• Accelerating renal clearance

Correct Answer: Stabilizing tertiary and quaternary structure through covalent crosslinks

Q17. Insulin lispro achieves a more rapid onset of action relative to regular human insulin primarily because of what modification?

• Fusion to an Fc fragment
• PEGylation of the A‑chain
• Reversal of amino acids at B28 and B29 reducing hexamer formation
• Addition of large glycan chains to increase solubility

Correct Answer: Reversal of amino acids at B28 and B29 reducing hexamer formation

Q18. Which analytical technique provides the most detailed information on molecular mass and glycoform heterogeneity of a recombinant protein?

• SDS‑PAGE with silver staining
• Enzyme‑linked immunosorbent assay (ELISA)
• LC‑MS (liquid chromatography–mass spectrometry)
• Dynamic light scattering (DLS)

Correct Answer: LC‑MS (liquid chromatography–mass spectrometry)

Q19. A common trade‑off observed with PEGylation of therapeutic proteins is:

• Increased receptor affinity with decreased half‑life
• Decreased immunogenicity but also reduced receptor binding affinity and potency
• Enhanced aggregation due to PEG hydrophobicity
• Elimination of the need for sterile filtration

Correct Answer: Decreased immunogenicity but also reduced receptor binding affinity and potency

Q20. Which excipient is commonly used to stabilize proteins during lyophilization and reduce aggregation on reconstitution?

• Polysorbate 80 exclusively (no sugars required)
• Sodium chloride as sole stabilizer
• Trehalose (a non‑reducing disaccharide) to protect structure during drying
• Ethanol to prevent microbial growth

Correct Answer: Trehalose (a non‑reducing disaccharide) to protect structure during drying

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