Development of a chromatography-free method for high-throughput MS-based bioanalysis of therapeutic monoclonal antibodies

• Main Authors: Li, N. (Author), Wang, S. (Author), Yan, Y. (Author), Zhang, Z. (Author)
• Format: Article
• Language: English
• Published: Future Medicine Ltd. 2021
• Subjects: Antibodies, Monoclonal; Article; controlled study; drug development; Drug Development; feasibility study; fractionation; high throughput analysis; high throughput screening; high-throughput bioanalysis; High-Throughput Screening Assays; human; Humans; immunoglobulin G1; immunoglobulin G3; immunoglobulin G4; intermethod comparison; LC-free; limit of quantitation; liquid chromatography; mass spectrometry; measurement accuracy; monoclonal antibody; multiplexed PRM; peptide derivative; proof of concept; quantitative analysis; sensitivity analysis; solid phase extraction; SPE; tandem mass spectrometry; Tandem Mass Spectrometry; therapeutic mAbs
• Online Access: View Fulltext in Publisher

 Aim: Our objective was to test the feasibility of developing an LC-free, MS-based approach for high-throughput bioanalysis of humanized therapeutic monoclonal antibodies. Methodology: A universal tryptic peptide from human IgG1, IgG3 and IgG4 was selected as the surrogate peptide for quantitation. After tryptic digestion, the surrogate peptide was fractionated via solid-phase extraction before being subjected to direct infusion-based MS/MS analysis. A high-resolution, multiplexed (MSX = 2) parallel reaction monitoring method was developed for data acquisition. Results & conclusion: This proof-of-concept study demonstrated the feasibility of achieving high-throughput MS-based bioanalysis of monoclonal antibodies using an LC-free workflow with sensitivity comparable to conventional LC-MS/MS-based methods. © 2021