Generic Hybrid Ligand Binding Assay Liquid Chromatography High-Resolution Mass Spectrometry-Based Workflow for Multiplexed Human Immunoglobulin G1 Quantification at the Intact Protein Level: Application to Preclinical Pharmacokinetic Studies

摘要: The quantitative analysis of human immunoglobulin G1 (hIgG1) by mass spectrometry is commonly performed using surrogate peptides after enzymatic digestion. Since some limitations are associated with this approach, a novel workflow presented hybridizing ligand binding assay (LBA) liquid chromatography–high-resolution (LC–HRMS) for hIgG1 quantification directly at the intact protein level. Different hIgG1s, including [13C]-labeled version used as internal standard, were immuno-enriched from rat serum fully automated platform based on streptavidin coated tips and biotinylated mouse anti-hIgG capture antibody targeting fragment crystallizable region followed overnight deglycosylation prior to LC–HRMS analysis. proposed utilized extracted ion chromatograms (XICs) nondeconvoluted full-scan MS spectrum. was validated in terms selectivity, sensitivity, accuracy/precision, carry-over, dilution linearity, reproducibil...