Mitigating ADA matrix interferences: semiquantitative double immunoprecipitation for anti-bevacizumab antibody in
Yu Gus Hui1, Fang Wang1, Jian Wu1
1Department of Chromatographic Services, PPD Laboratory Services (a Part of Thermo Fisher Scientific), Henrico, VA, USA.
Background:
Hybrid LC - MS/MS methodology offers orthogonal ADA quantitation but is limited by endogenous immunoglobulin matrix interference challenges.
Methods:
We developed a novel semi-quantitative LC-MS/MS isotyping assay for rabbit IgG/IgM immunogenicity in K2EDTA plasma using double immunoprecipitation with double acid dissociation.
Results And Conclusions:
As compared to single immunoprecipitation, double immunoprecipitation reduced endogenous IgG by ≥90% and IgM by >95% (W1B1) or >55% (W3B2), improved peak-area ratios, and achieved an LLOQ of 100 ng/mL, with acceptable accuracy and precision. Double immunoprecipitation plus acid dissociation mitigates matrix interference and enables robust semi-quantitative hybrid LC-MS/MS ADA isotyping in rabbit plasma.
