Characterization of an anti-human ovarian carcinomaxanti-human CD3 bispecific single-chain antibody with an albumin-original interlinker.
OBJECTIVE
The objective of this study was to determine the properties of a single-chain bispecific antibody (scBsAb) against human ovarian carcinoma and to develop this agent for potential use in human ovarian cancer.


METHODS
ELISA and FACS were performed to determine the antigen-binding properties of the scBsAb. Its abilities to retarget the pre-activated human peripheral blood mononuclear cells (PBMCs) to human ovarian carcinoma cell line SKOV3 cells and mediate their lysis in vitro were performed by a colorimetric MTT-based assay. Nude mice bearing human SKOV3 tumor xenografts were used to study the distribution and imaging of the scBsAb. Its pharmacokinetics in vivo was also studied in naive BALB/c mice.


RESULTS
The scBsAb showed nearly identical ligand binding properties at each site relative to the individual monovalent single-chain antibody prototype molecules and could bridge SKOV3 and human T cell line Jurkat, which expresses CD3 antigens on the surface of cells together. It can also retarget the pre-activated PBMCs to SKOV3 cells and mediated their lysis in vitro effectively. Imaging and distribution study demonstrated that the antibody could target the tumor. Its elimination in vivo corresponded to second-order kinetics with a terminal half-life time (t(1/2)beta) of 7.7 h.


CONCLUSIONS
This scBsAb with easy production and reasonable blood retention time should be developed for potential use in human ovarian cancer.
Fang
Min
M
Group 102, Institute of Genetics and Developmental Biology, Academia Sinica, Beijing 100101, China.
Zhao
Rui
R
Yang
Zhi
Z
Zhang
Zhong
Z
Li
Hua
H
Zhang
Xue-Tao
XT
Lin
Qing
Q
Huang
Hua-Liang
HL
D000418:Albumins / Q000737:chemistry / Q000493:pharmacokinetics; D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology* / Q000493:pharmacokinetics / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology*; D045744:Cell Line, Tumor; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000493:pharmacokinetics; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D018796:Immunoconjugates / Q000276:immunology / Q000493:pharmacokinetics; D019169:Jurkat Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D010051:Ovarian Neoplasms / Q000000981:diagnostic imaging / Q000276:immunology* / Q000378:metabolism; D011877:Radionuclide Imaging; D013667:Technetium / Q000493:pharmacokinetics; D014018:Tissue Distribution
D000418:Albumins; D018033:Antibodies, Bispecific; D017252:CD3 Complex; D003432:Cross-Linking Reagents; D018796:Immunoconjugates; D013667:Technetium