Calculated conjugated payload from immunoassay and LC-MS intact protein analysis measurements of antibody-drug conjugate.
OBJECTIVE
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.


CONCLUSIONS
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.
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Brian
B
Rago
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Tracey
T
Clark
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Lindsay
L
King
Clinical Assay Group, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Jenny
J
Zhang
World Wide Medicinal Chemistry, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
L Nathan
LN
Tumey
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Fengping
F
Li
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Frank
F
Barletta
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Cong
C
Wei
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Mauricio
M
Leal
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Steve
S
Hansel
Pharmacokinetics, Dynamics & Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.|Present affiliation: PKDM Amgen, 360 Binney Street, Cambridge, MA, USA.
Xiaogang
X
Han
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D006207:Half-Life; D007118:Immunoassay; D018796:Immunoconjugates / Q000097:blood* / Q000493:pharmacokinetics; D013058:Mass Spectrometry; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D000068878:Trastuzumab / Q000737:chemistry
D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D004364:Pharmaceutical Preparations; D000068878:Trastuzumab
LC–MS; PKs; antibody-drug conjugate; ligand-binding assay; metabolism