DATA INDEPENDENT ACQUISITION MODES FOR IDENTIFICATION, QUANTIFICATION AND MONITORING OF LOW-ABUNDANCE HOST CELL PROTEINS DURING MONOCLONAL ANTIBODY BIOPROCESSING

摘要

Quantification of large molecular weight biotherapeutics in biological matrices relies mostly on enzymelinked immunosorbent assays (ELISAs) mainly due to several advantages: good sensitivity (1-10 ppm), highthroughput, ease-of-use and low cost per sample. In the case of low-abundance protein impurities (HCPs) present in protein therapeutics (e.g. mAbs), these biological assays typically provide the total HCP concentration (usually expressed in ppm or ng HCP/mg mAb), but ELISAs cannot identify and measure individual HCP contaminants. Several MS-based assays have been recently developed to cover this knowledge gap [1-2]. Here we explored the capabilities of a single dimension chromatographic assay, coupled with mass spectrometry using two different data independent acquisition modes for detection of peptide precursors and their fragments.