The Application of Capability Indices in The Validation of ELISA Methodology

Abstract

Aim: To investigate the impact of sample replication number (duplicate vs triplicate) on the validation of ELISA methodology. Materials & methods: The methodology was validated with reference sample and test sample as an 11-point triplicate dilution series. The data were reanalyzed post-validation as if conducted as a duplicate dilution series. Results: The triplicate methodology was validated with a precision of 5.3% and mean bias of -1.7%. The duplicate methodology generated a precision of 5.7% and mean bias of -2.2%. Conclusion: Both the triplicate (method capability index = 1.37) and duplicate (method capability index = 1.25) ELISA methodology can support an 80–125% relative potency specification with a 0.004% or 0.018% probability of out-of-specification results, respectively.

Keywords:

• antibody–drug conjugate
• ELISA
• parallel curve analysis (parallelism)
• potency
• relative potency

Financial disclosure

The author has no competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Competing interests disclosure

The data presented are derived from original work conducted at Piramal Pharma Solutions paid for by an external client. The studies were conducted using concepts detailed in US Pharmacopeia monographs, and the client gave permission to create a dataset, so that examples could be presented in this article. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.