Bioanalysis of protein drugs, soluble targets, and biomarkers, presents many challenges. Biotherapeutic modalities are increasing in complexity with many multi-domain molecular forms, including fusion proteins, bispecifics and ADCs. As complementary or alternative approaches to traditional ligand-binding assays (LBAs), hybrid LBA/LC–MS techniques combine the advantages of immunoaffinity capture for selective and efficient enrichment of protein analytes (and/or on-line purification of target peptides) with the benefits of “direct” physicochemical-based detection offered by LC–MS technologies. This presentation introduced the variety of hybrid LBA/LC–MS approaches that are available and discussed some practical aspects in choosing a format, optimizing and evaluating assay performance.
What will you learn?
- Differences and practical considerations in choosing a physical format and critical reagent type for the LB “front end” of a hybrid method
- Understanding LBA-related issues that may also impact a hybrid method and how to mitigate and evaluate them
- Examples illustrating the value of generic approaches to save time and cost vs. highly specific targeted approaches to maximize sensitivity
Who may this interest?
- CROs, Pharma and Biotech companies focused on LC–MS/MS quantitative bioanalysis of large molecules
- Mass spectrometrists and bioanalytical chemists
- Academicians and students interested in LC–MS/MS bioanalysis
Rand Jenkins is Scientific Director for the Chromatographic Sciences Department (CSD) of PPD Laboratories bioanalytical lab in Richmond (VA, USA) and Middleton (WI, USA). Since obtaining a BS in chemistry from the University of Nevada Reno in 1972, Rand has been involved for over 40 years in the development and applications of GC– and LC–MS technologies in the environmental and pharmaceutical fields. He joined PPD in 1994 and currently provides scientific guidance to the R&D teams in bioanalytical methods development and validation. Rand’s major focus in recent years has been to establish and promote bioanalysis of peptides and proteins using LC–MS technology.