Direct Bioanalysis of ADCs using Affinity Capture-LC-HR/AMS Techniques for Characterization ... PDF
Preview Direct Bioanalysis of ADCs using Affinity Capture-LC-HR/AMS Techniques for Characterization ...
Direct Bioanalysis of ADCs using AAffffiinniittyy CCaappttuurree--LLCC--HHRRAAMMSS Techniques for Characterization and Quantification—a Progress UUppddaattee Rand Jenkins et al EBF 8th Open Symposium BBaarcceelloonnaa SSppaaiinn, 1188 NNoov 22001155 Overview 1 Biotherapeutics diversity 22 Whhy new BA approachhes are needdedd 33 LLCC-MMSS pprrootteeiinn bbiiooaannaallyyssiiss wwoorrkkfflloowwss 44 AADDCC cchhaarraacctteerriizzaattiioonn 55 IInnttaacctt AADDCC qquuaannttiiffiiccaattiioonn 6 Oppen qquestions & final thougghts 2 Biotherappeutics diversityy ppeeppttiiddeess mAbs bispecifics (bsAbs, DART®s, BiTE®s) AADDCCss prodrug/carrier conjugates (albumin, PEG) ffusiion protteiins messenger RNA therapeutics™ oligonucleotides (ssDNA, siRNA, mRNA) CCAARRss 3 Alternative formats for bispecifics CChhriisttophhSSpiiess, QQiianttiingZZhhaii, PPaull JJ. CCartter, AAlltternattiive mollecullar fformatts andd tthherapeuttiic applications for bispecific antibodies,Molecular Immunology 67 (2015) 95–106 4 Antibodyy-Drugg Conjjuggates KKaur SS et all. BBiioanallysiis, ((22001133)) 55((22)), 220011–222266 5 GlyycoConnect ADC: a new gglyycan conjjuggation idea Remon van Geel et al, Chemoenzyymatic Conjjuggation of Toxic Payyloads to the Globallyy Conserved N-Glyycan of Native mAbs Provides Homogeneous and Highly Efficacious Antibody−Drug Conjugates, BioconjugateChem.2015, in press 6 Whyy new BA apppproaches are needed ++ BBiiootthheerraappeeuuttiiccss iinnnnoovvaattiioonn iiss pprroodduucciinngg iinnccrreeaassiinnggllyy ccoommpplleexx molecular constructs + Their inherent heterogeneity and potential to undergo stability- and/or catabolism-related changes are driving the need for greater characterization ++ CCoonnvveennttiioonnaall LLiiggaanndd BBiinnddiinngg AAssssaayyss ((LLBBAA)) aarree lliimmiitteedd iinn tthheeiirr ability to provide data reflecting molecular structure (e.g. Drugg Antibodyy Ratio ((DAR)) distribution for ADCs)) + MS-based methods, including affinity capture-liquid chhromattographhy-hhiighh resolluttiion accuratte mass specttromettry (AC-LC-HRAMS), are emerging as versatile tools 7 LLCC-MMSS pprrootteeiinn bbiiooaannaallyyssiiss wwoorrkkfflloowwss 8 Protein LC-MS//MS Bioanalyysis General Strateggyy Protein/Peptide in Biomatrix Sample MW < ~10 kDa MW > ~10 kDa Direct Measurement Indirect Measurement ((IInnttaacctt AAnnaallyyttee)) ((PPrrootteeoollyyttiicc PPeeppttiiddeess)) Extraction//Enrichment Extraction//Enrichment (Chemical (e.g. PPT, SPE) (Chemical (e.g. PPT) or Affinity Capture) or Affinity Capture) Clean up Proteolytic LC‐MS/MS (SPE or DDiiggeessttiioonn Peptide AC) LC‐MS/MS 9 Protein LC-HRAMS Bioanalyysis Alternative Strateggyy Protein/Peptide in Biomatrix Sample MW < ??? kDa MW > ~10 kDa Direct Measurement Indirect Measurement ((IInnttaacctt AAnnaallyyttee)) ((PPrrootteeoollyyttiicc PPeeppttiiddeess)) Enrichment Extraction//Enrichment Affinity Capture (Chemical (e.g. PPT) or Affinity Capture) LC‐HRAMS Clean up Proteolytic (SPE or DDiiggeessttiioonn Peptide AC) LC‐HRAMS 10
Description:The list of books you might like
