High-Throughput Mass Spectrometry for Biopharma: A Universal Modality and Target Independent Analytical Method for Accurate Biomolecule Characterization.
Iain D G CampuzanoEmma M Pelegri-O'DayNithya SrinivasanJennifer L LippensPascal EgeaAiko UmedaJennifer AralTianqi ZhangArthur D LaganowskyChawita NetirojjanakulPublished in: Journal of the American Society for Mass Spectrometry (2022)
Reversed-phase liquid chromatographic mass spectrometry (rpLC-MS) is a universal, platformed, and essential analytical technique within pharmaceutical and biopharmaceutical research. Typical rpLC method gradient times can range from 5 to 20 min. As monoclonal antibody (mAb) therapies continue to evolve and bispecific antibodies (BsAbs) become more established, research stage engineering panels will clearly evolve in size. Therefore, high-throughput (HT) MS and automated deconvolution methods are key for success. Additionally, newer therapeutics such as bispecific T-cell engagers and nucleic acid-based modalities will also require MS characterization. Herein, we present a modality and target agnostic HT solid-phase extraction (SPE) MS method that affords the analysis of a 96-well plate in 41.4 min, compared to the traditional rpLC-MS method that would typically take 14.4 h. The described method can accurately determine the molecular weights for monodispersed and highly polydispersed biotherapeutic species and membrane proteins; determine levels of glycosylation, glycation, and formylation; detect levels of chain mispairing; and determine accurate drug-to-antibody ratio values.
Keyphrases
- mass spectrometry
- liquid chromatography
- high throughput
- ms ms
- solid phase extraction
- high performance liquid chromatography
- gas chromatography
- multiple sclerosis
- high resolution
- monoclonal antibody
- high resolution mass spectrometry
- tandem mass spectrometry
- capillary electrophoresis
- simultaneous determination
- nucleic acid
- machine learning
- single cell
- gas chromatography mass spectrometry
- single molecule