A rapid and quantitative technique for assessing IgG monomeric purity, calibrated with the NISTmAb reference material

• DOI: 10.1007/s00216-019-02029-0
• Publication Date: 2019-08-02T12:04:25Z
• Authors: Peter P. Reader; Rouslan V. Olkhov; Shaun Reeksting; Anneke Lubben; Christopher J. Hyde; Andrew M. Shaw
• Keywords: Avidity

The fraction of intact monomer in a sample (moles/moles), the monomeric purity, is measured as quality control therapeutic monoclonal antibodies but often unknown research samples and remains major source variation quantitative antibody-based techniques such immunoassay development. Here, we describe novel multiplex technique for estimating purity antigen affinity grade antibody samples. Light scattering was used to simultaneously observe mass binding biosensor surfaces functionalised with (revealing Fab kinetics) or protein A/G (PAG). Initial estimates 7 including infliximab biosimilar were estimated by observing deficit on PAG surface compared NISTmAb standard high purity. Monomeric improved second step surface. displayed tightly controlled stoichiometries 1.31 ± 0.57 1.71 0.16 (95% confidence interval)-within theoretical limit 1-2 antigens per depending avidity. other panel range 0.06-1.15, attributed lower verified electrospray ionization spectrometry (ESI), gold structural characterization antibodies. ESI data indicated that had values 93.5% 94.7%, respectively, whilst significantly (54-89%). Our results demonstrate rapid testing (< 15 min) which could improve reproducibility experiments.