Antibody-drug conjugate bioanalysis using LB-LC-MS/MS hybrid assays: strategies, methodology and correlation to ligand-binding assays.
BACKGROUND
Antibody-drug conjugates (ADCs) are complex drug constructs with multiple species in the heterogeneous mixture that contribute to their efficacy and toxicity. The bioanalysis of ADCs involves multiple assays and analytical platforms.


METHODS
A series of ligand binding and LC-MS/MS (LB-LC-MS/MS) hybrid assays, through different combinations of anti-idiotype (anti-Id), anti-payload, or generic capture reagents, and cathepsin-B or trypsin enzyme digestion, were developed and evaluated for the analysis of conjugated-payload as well as for species traditionally measured by ligand-binding assays, total-antibody and conjugated-antibody.


CONCLUSIONS
Hybrid assays are complementary or viable alternatives to ligand-binding assay for ADC bioanalysis and PK/PD modeling. The fit-for-purpose choice of analytes, assays and platforms and an integrated strategy from Discovery to Development for ADC PK and bioanalysis are recommended.
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Jian
J
Wang
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Huidong
H
Gu
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Ang
A
Liu
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Alexander
A
Kozhich
Biologics Discovery California, Bristol-Myers Squibb, Redwood City, CA 95035, USA.
Vangipuram
V
Rangan
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Heather
H
Myler
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Linlin
L
Luo
Preclinical Candidate Optimization, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Richard
R
Wong
Preclinical Candidate Optimization, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Huadong
H
Sun
Drug Safety Evaluation, Bristol-Myers Squibb, New Brunswick, NJ 08903, USA.
Bonnie
B
Wang
Clinical Pharmacology & Pharmacometrics, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Heather E
HE
Vezina
Biologics Discovery California, Bristol-Myers Squibb, Redwood City, CA 95035, USA.
Shrikant
S
Deshpande
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Yan
Y
Zhang
Preclinical Candidate Optimization, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Zheng
Z
Yang
Preclinical Candidate Optimization, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Timothy V
TV
Olah
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Anne-Francoise
AF
Aubry
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Mark E
ME
Arnold
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Renuka
R
Pillutla
Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Binodh
B
DeSilva
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D000882:Haplorhini; D006801:Humans; D007118:Immunoassay / Q000379:methods; D018796:Immunoconjugates / Q000032:analysis / Q000097:blood*; D057230:Limit of Detection; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000097:blood*; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods*
D018796:Immunoconjugates; D004364:Pharmaceutical Preparations
DAR-bias; DAR-sensitive ADC assay; antibody conjugated-payload; antibody–drug conjugate; drug antibody ratio; fit-for-purpose bioanalysis; generic reagents; immunocapture LC–MS/MS hybrid assay; ligand binding-LC–MS/MS hybrid assay; soluble target