Interlaboratory Studies Using the NISTmAb to Advance Biopharmaceutical Structural Analytics

Abstract

Biopharmaceuticals such as monoclonal antibodies are required to be rigorously characterized using a wide range of analytical methods. Various material properties must be characterized and well controlled to assure that clinically relevant features and critical quality attributes are maintained. A thorough understanding of analytical method performance metrics, particularly emerging methods designed to address measurement gaps, is required to assure methods are appropriate for their intended use in assuring drug safety, stability, and functional activity. To this end, a series of interlaboratory studies have been conducted using NISTmAb, a biopharmaceutical-representative and publicly available monoclonal antibody test material, to report on state-of-the-art method performance, harmonize best practices, and inform on potential gaps in the analytical measurement infrastructure. Reported here is a summary of the study designs, results, and future perspectives revealed from these interlaboratory studies which focused on primary structure, post-translational modifications, and higher order structure measurements currently employed during biopharmaceutical development.

2D NMR Analysis as a Sensitive Tool for Evaluating the Higher-Order Structural Integrity of Monoclonal Antibody against COVID-19

“…The CCSD analysis ( Figure 2 ) also indicated that batch-to-batch variability was small, as the average CCSD value was 5.2 ± 2.5 ppb at T0. Our CCSD analysis is consistent with the NMR data already reported on monoclonal antibodies to assess their higher order structures [ 10 , 17 ].…”

Comparison of N-Glycopeptide to Released N-Glycan Abundances and the Influence of Glycopeptide Mass and Charge States on N-Linked Glycosylation of IgG Antibodies

“…Except for in-source generated glycopeptides, the findings on NISTmAb glycosylation interlaboratory studies 45,46 on released N-glycans are consistent, with the relative GADS and FLD glycoform abundances of G4H3F, G4H4F, and G4H5F accounting for 85% of the total (Figures 2−4). GADS values seem to provide reasonable estimates of the relative concentrations at the protein level.…”

Advances in Mass Spectrometry‐Based Structural Proteomics: Development of HDX‐MS and XL‐MS Techniques from Recombinant Protein to Cellular Systems

“…In other words, alterations in backbone amide hydrogen chemical environment such as changes in hydrogen bonding and/or solvent accessibility result in measurable changes in HDX rates. Applications for continuous labeling HDX‐MS are far reaching in both academic and industry settings and have been used to map protein‐protein interactions (PPIs) or protein interactions with cognate ligand, small molecules, nucleic acids, and antibodies [27–30] …”

“…Applications for continuous labeling HDX-MS are far reaching in both academic and industry settings and have been used to map protein-protein interactions (PPIs) or protein interactions with cognate ligand, small molecules, nucleic acids, and antibodies. [27][28][29][30] Alternatively, HDX-MS experiments can also utilize a pulsed labeling approach (Figure 2C -pulsed labeling). [31] In this workflow protein samples are first perturbed for different time intervals and then subjected to a single, short deuterium labeling step.…”