Hydrolytically Stable Site-Specific Conjugation at the N-Terminus of an Engineered Antibody.
Antibody-drug conjugates (ADCs) have emerged as an important class of therapeutics for cancer treatment that combine the target specificity of antibodies with the killing activity of anticancer chemotherapeutics. Early conjugation technologies relied upon random conjugation to either lysine or cysteine residues, resulting in heterogeneous ADCs. Recent technology advancements have resulted in the preparation of homogeneous ADCs through the site-specific conjugation at engineered cysteines, glycosylated amino acids, and bioorthogonal unnatural amino acids. Here we describe for the first time the conjugation of an anti-mitotic drug to an antibody following the mild and selective oxidation of a serine residue engineered at the N-terminus of the light chain. Using an alkoxyamine-derivatized monomethyl auristatine E payload, we have prepared a hydrolytically stable ADC that retains binding to its antigen and displays potent in vitro cytotoxicity and in vivo tumor growth inhibition.
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Pamela
P
Thompson
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Binyam
B
Bezabeh
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Ryan
R
Fleming
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Monica
M
Pruitt
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Shenlan
S
Mao
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Patrick
P
Strout
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Cui
C
Chen
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Song
S
Cho
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Haihong
H
Zhong
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Herren
H
Wu
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Changshou
C
Gao
Antibody Discovery and Protein Engineering, ‡Oncology Research, MedImmune , Gaithersburg, Maryland 20878, United States.
Nazzareno
N
Dimasi
D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D008819:Mice, Nude; D010091:Oximes / Q000737:chemistry; D015202:Protein Engineering / Q000379:methods*; D055550:Protein Stability; D051381:Rats; D036104:Receptor, EphA2 / Q000276:immunology / Q000378:metabolism; D012694:Serine / Q000737:chemistry; D023041:Xenograft Model Antitumor Assays
D000906:Antibodies; D000970:Antineoplastic Agents; D010091:Oximes; D012694:Serine; D036104:Receptor, EphA2