title: Straightforward Glycoengineering Approach to Site-Specific Antibody-Pyrrolobenzodiazepine Conjugates.
abstract: Antibody-drug conjugates (ADCs) have become a powerful platform to deliver cytotoxic agents selectively to cancer cells. ADCs have traditionally been prepared by stochastic conjugation of a cytotoxic drug using an antibody's native cysteine or lysine residues. Through strategic selection of the mammalian expression host, we were able to introduce azide-functionalized glycans onto a homogeneously glycosylated anti-EphA2 monoclonal antibody in one step. Conjugation with an alkyne-bearing pyrrolobenzodiazepine dimer payload (SG3364) using copper-catalyzed click chemistry yielded a site-specific ADC with a drug-to-antibody ratio (DAR) of four. This ADC was compared with a glycoengineered DAR two site-specific ADC, and both were found to be highly potent against EphA2-positive human prostate cancer cells in both anin vitrocytotoxicity assay and a murine tumor xenograft model.
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Pamela
authors.initials: P
authors.lastname: Thompson
authors.affiliation: QMB Innovation Centre, Spirogen , 42 New Road, E1 2AX London, U.K.
authors.forename: Ebele
authors.initials: E
authors.lastname: Ezeadi
authors.affiliation: QMB Innovation Centre, Spirogen , 42 New Road, E1 2AX London, U.K.
authors.forename: Ian
authors.initials: I
authors.lastname: Hutchinson
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Ryan
authors.initials: R
authors.lastname: Fleming
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Binyam
authors.initials: B
authors.lastname: Bezabeh
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Jia
authors.initials: J
authors.lastname: Lin
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Shenlan
authors.initials: S
authors.lastname: Mao
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Cui
authors.initials: C
authors.lastname: Chen
authors.affiliation: QMB Innovation Centre, Spirogen , 42 New Road, E1 2AX London, U.K.
authors.forename: Luke
authors.initials: L
authors.lastname: Masterson
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Haihong
authors.initials: H
authors.lastname: Zhong
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Dorin
authors.initials: D
authors.lastname: Toader
authors.affiliation: QMB Innovation Centre, Spirogen , 42 New Road, E1 2AX London, U.K.
authors.forename: Philip
authors.initials: P
authors.lastname: Howard
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Herren
authors.initials: H
authors.lastname: Wu
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Changshou
authors.initials: C
authors.lastname: Gao
authors.affiliation: Antibody Discovery and Protein Engineering and Oncology Research, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Dimasi
keywords: Antibody−drug conjugates; SG3364; carbohydrate remodeling; click chemistry; pyrrolobenzodiazepine; site-specific conjugation