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Q&A: Which are the advantages and limitations of orthogonal methods for HCP monitoring?

Q & A

Answer: There is an increased need for simultaneous HCP detection and identification to improve HCP risk assessment.

This can be achieved using antibody-dependent methods such as the ELISA, in combination with orthogonal LC-MS analyses. The ELISA remains the Gold Standard for HCP monitoring, however LC-MS can provide complementary information. The ELISA is easy to implement, suitable for high-throughput measurements and accepted for the quantification of HCP residues by regulatory agencies for the approval of drug release. LC-MS analyses can be used as an additional method to validate the HCP ELISA performance for a specific manufacturing process, while enabling the identification of single HCPs to assess their individual safety risk. In the following the strengths and drawbacks of each method are specified:

 HCP-ELISALC-MS
Strengths     
  • Easy to implement
  • Continuous monitoring possible
  • High-throughput method
  • Easy to transfer into GMP-environment
  • Highly sensitive, even with high drug substance concentrations
  • Relative Assay robustness
  • Strong signal of highly immunological antigens aids subsequent risk assessment
  • High precision and accuracy
  • Antibody independent -> fast implementation
  • Batch comparability
  • Identification and quantification of HCPs
  • Detection of HCPs not covered by antibodies
Drawbacks 
  • Antibody dependent -> generation is time-consuming
  • Total HCP signal only, no information about identity and concentration of individual HCPs
  • Unable to detect HCPs that do not react with ELISA antibodies
  • Absolute quantification requires precise standards
  • Current technology does not match the ELISA sensitivity/ detection limit for HCP profiling in final drug
  • Requires expensive and sophisticated equipment + high-level hands-on experience
  • No USP standard established  method transfer into GMP-environment difficult
  • Underrepresentation of detectable HCPs which are resistant to tryptic digest or result in bad ionization
  • Not suitable for high-throughput

Considering all strengths and drawbacks of both, HCP-ELISA and LC-MS methods, optimal HCP moitoring is achieved by combining their advantages:

  • Antibody-based methods and MS-only techniques rely on completely different methodological approaches and thus provide true orthogonality
  • The combination of both methods enables a solid HCP monitoring strategy based on ELISA technology including quantification and identification of relevant HCPs
  • Optimal risk assessment together with high-sensitive HCP detection: supports the drug safety profile, increases chances for successful Market Authorization Application (MAA) and benefits patient safety

Do you want to discuss HCP monitoring and the advantages of orthogonal methods in further detail with one of our experts? Let's make an appointment.

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