title: Fusion of chemotactic peptide to a single-chain bi-specific antibody (scBsAb) potentiates its cytotoxicity to target tumour cells.
abstract: Anti-tumour BsAb (bi-specific antibody) has been proved very effective in killing tumour cells both in vitro and in vivo. In order to enhance its ability to recruit and activate T-lymphocytes and then improve tumour-specific cytolysis, an anti-ovarian carcinoma/CD3 BsAb, BHL-I, was fused to N-terminal 18 peptide of CCL21 (CC chemokine ligand 21) to produce a new chemotactic BsAb, named 18TBHL. It was expressed in soluble form in the cytoplasm of Escherichia coli and purified with DEAE anion-exchange chromatography and immobilized-metal-ion affinity chromatography. The chemotactic ability of 18TBHL to PBLs (peripheral-blood lymphocytes) was detected by Boyden chamber chemotaxis assay. The specific ability to bind to ovarian carcinoma cells, SKOV3, and PBLs was tested by ELISA and flow cytometry. Just as expected, the enhanced tumour-specific cytolysis of 18TBHL was validated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] method and flow cytometry. The results indicated that fusion of chemotactic peptide to BsAb potentiated its cytotoxicity to tumour cells in vitro. It suggests that 18TBHL may be a promising candidate agent in cancer immunotherapy.
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, Beijing 100101, People's Republic of China.
authors.forename: Jingzhen
authors.initials: J
authors.lastname: Song
authors.forename: Xiangbin
authors.initials: X
authors.lastname: Wang
authors.forename: Chun
authors.initials: C
authors.lastname: Lei
authors.forename: Jinhua
authors.initials: J
authors.lastname: Piao
authors.forename: Changcheng
authors.initials: C
authors.lastname: Yin
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zhang
authors.forename: Qing
authors.initials: Q
authors.lastname: Lin
authors.forename: Hualiang
authors.initials: H
authors.lastname: Huang
mesh_terms: D018033:Antibodies, Bispecific / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D017209:Apoptosis; D054421:Chemokine CCL21; D019742:Chemokines, CC / Q000737:chemistry*; D002633:Chemotaxis / Q000187:drug effects / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D016503:Drug Delivery Systems / Q000379:methods; D004357:Drug Synergism; D005260:Female; D006801:Humans; D050576:Membrane Fusion Proteins / Q000276:immunology / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000276:immunology* / Q000628:therapy*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D014407:Tumor Cells, Cultured
chemical_list: D018033:Antibodies, Bispecific; C515478:CCL21 protein, human; D054421:Chemokine CCL21; D019742:Chemokines, CC; D050576:Membrane Fusion Proteins; D010446:Peptide Fragments