LC/MSversus Immune‐Based Bioanalytical Methods in Quantitation of Therapeutic Biologics in Biological Matrices

Abstract

This chapter reviews the main characteristics of liquid chromatography/mass spectrometry (LC/MS) and ligand‐binding assay (LBA) methods. If an LBA method for biotherapeutics quantification is well established, it usually provides sufficient sensitivity and selectivity and is considered the method of choice. The LC/MS method emerged as an alternative strategy for quantification of proteins of interest in complex biological matrices. Multiplexed capacity and selectivity are critical features determining the applicability of a bioanalytical method. Well‐established LBA methods usually provide adequate sensitivity for quantification analysis of biotherapeutics in preclinical/clinical pharmacokinetic (PK)/pharmacodynamic (PD) study. LC/MS with high resolution analyzer has become a promising technology for characterization of antibody‐drug conjugate (ADC) in the recent years. An affinity capture capillary LC coupled to quadrupole‐time‐of‐flight (TOF) MS has been employed to analyze anti‐MUC16 TDC, which obtained both in vitro and in vivo drug‐to‐antibody ratios (DARs) information.

Understanding the pharmacokinetic journey of Fc-fusion protein, rhIL-7-hyFc using complementary approach of two analytical methods, accelerator mass spectrometry and ELISA

“…However, its specificity and accuracy can be reduced if the analytes of interest are modified or degraded, which can be further complicated by interference by ADA. The performance of the ELISA can be also affected by matrix effects and inter-species differences [ 10 , 11 ]. AMS is an emerging quantitative tool, which is useful not only for chemical compounds but also for protein pharmaceuticals in early drug development due to its high sensitivity [ 12–14 ].…”

Challenges of non-clinical safety testing for biologics: A Report of the 9th BioSafe European Annual General Membership Meeting

“…55 LC-MS is gaining popularity for being highly selective and possessing multiplexing ability, whereas LBA performs better in terms of throughput and sensitivity. [56][57][58][59] A decision-tree that helps to select the right assay and three case studies using this approach were presented. Some factors to considers are: limit of quantification to be achieved, moieties to be quantified (total drug vs free drug) and whether in vivo integrity of the compound must be confirmed.…”