In this interview, John Kellie (GSK, PA, USA) discusses the technologies he uses on a regular basis, touching on how intact mass analysis and whole molecule protein quantification using LC–MS can provide additional information to make more informed decisions in the preclinical/clinical space. He explains how large molecule bioanalysis directly influences downstream drug development and his future outlook on where the field will be in 5–10 years’ time.
In this application note, Yun Alelyunas (Waters Corporation) describes an LC-HRMS method for high sensitivity bioanalysis of monoclonal antibodies by direct mass measurement at the intact level for whole molecule quantification. This emerging approach can serve as an alternative or complement to traditional methods such as ligand binding assay for protein quantification. Methodology and guidance for evaluating and optimizing data processing parameters are discussed in detail.
Whole molecule quantification of protein therapeutics by LC-HRMS is emerging as a complement to the established technique of protein quantification by LC-MS/MS using surrogate peptides. In this publication, John Kellie (GSK) discusses the applicability of HRMS for quantification of intact proteins and subunits, and presents guidance for processing data to quantify biotherapeutics in the range of 12-25 kDa from biological matrices.