Calculated conjugated payload from immunoassay and LC–MS intact protein analysis measurements of antibody-drug conjugate. (November 2016)

• Record Type: Journal Article
• Authors: Rago, Brian; Clark, Tracey; King, Lindsay; Zhang, Jenny; Tumey, L Nathan; Li, Fengping; Barletta, Frank; Wei, Cong; Leal, Mauricio; Hansel, Steve; Han, Xiaogang
• Is Part Of: Bioanalysis. Volume 8:Number 21(2016)
• Page Start: 2205
• Page End: 2217
• Publication Date: 2016-11
• Subjects: antibody-drug conjugate -- LC–MS -- ligand-binding assay -- metabolism -- PKs; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2016-0160
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: Complex nature of bioconjugates require multiple bioanalytical approaches to support PK and absorption, distribution, metabolism and excretion characterization. For antibody-drug conjugate (ADC) bioanalysis both LC–MS and ligand-binding assays (LBAs) are employed.Results: A method consisting of immunocapture extraction of ADC from biomatrices followed by LC–MS analysis of light and heavy chain is described. Drug antibody ratio (DAR) profiles of ADC Tras-mcVC-PF06380101 dosed at 0.3, 1 and 3 mg/kg in Sprague Dawley rats were obtained. Combined with total antibody (monoclonal antibody) measurement by LBA, conjugated payload concentration was calculated.Conclusion: PK profiles from LBA, ADC and calculated conjugated payload (DAR × monoclonal antibody) were in good agreement. We present a new tool for PK assessment of ADCs while also exploring ADC metabolism and DAR sensitivity of LBA ADC assay.