Improvement of tumor targeting and antitumor activity by a disulphide bond stabilized diabody expressed in Escherichia coli.
We have generated an anti-Pgp/anti-CD3 diabody which can effectively inhibit the growth of multidrug-resistant human tumors. However, the two chains of the diabody are associated non-covalently and are therefore capable of dissociation. Cysteine residues were introduced into the V-domains to promote disulphide cross-linking of the dimer as secreted by Escherichia coli. Compared with the parent diabody, the ds-Diabody obtained was more stable in human serum at 37 degrees C, without loss of affinity or cytotoxicity activity in vitro. Furthermore, the ds-Diabody showed improved tumor localization and a twofold improved antitumor activity over the parent diabody in nude mice bearing Pgp-overexpressing K562/A02 xenografts. Our data demonstrate that ds-Diabody may be more useful in therapeutic applications than the parent diabody.
Liu
Juanni
J
State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, 300020 Tianjin, People's Republic of China. liujuanni@gmail.com
Yang
Ming
M
Wang
Jinhong
J
Xu
Yuanfu
Y
Wang
Yan
Y
Shao
Xiaofeng
X
Yang
Chunzheng
C
Gao
Yingdai
Y
Xiong
Dongsheng
D
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000276:immunology*; D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use*; D017252:CD3 Complex / Q000276:immunology*; D002460:Cell Line; D004220:Disulfides / Q000737:chemistry; D004355:Drug Stability; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D006801:Humans; D020014:K562 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000628:therapy*; D023041:Xenograft Model Antitumor Assays
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018033:Antibodies, Bispecific; D017252:CD3 Complex; D004220:Disulfides