title: Expression and purification of single-chain anti-HBx antibody in Escherichia coli.
abstract: Monoclonal antibodies have been widely used in tumor targeting studies with promising results. However, their clinical application has been limited by heterogeneity and macro-molecular movement of murine antibody. In this study, the variable-region (heavy- and light-chain) fragments of anti-HBx monoclonal antibody were enriched by the polymerase chain reaction. The expression vector, which included a 6x histidine sequence in the 3' terminus of the HBx single-chain antibody (sFv) was recombined with a linker sequence (KLGGGGFSGA) between the variable regions. The expression product from Escherichia coli fused with 6xHis was purified by nickel (Ni2+) nitrilotriacetate chelating resin. The results of enzyme-linked immunosorbent assay and Western blotting showed that sFv had binding affinity with HBxAg, suggesting that it could become a novel targeting carrier in clinical trials.
authors.affiliation: Liver Cancer Institute, Shanghai Medical University, PR China.
authors.forename: G
authors.initials: G
authors.lastname: Zhou
authors.forename: K D
authors.initials: KD
authors.lastname: Liu
authors.forename: H C
authors.initials: HC
authors.lastname: Sun
authors.forename: Y H
authors.initials: YH
authors.lastname: Chen
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Tang
authors.forename: C H
authors.initials: CH
authors.lastname: Schröder
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001483:Base Sequence; D015153:Blotting, Western; D004926:Escherichia coli / Q000378:metabolism*; D006511:Hepatitis B Antigens / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015534:Trans-Activators / Q000276:immunology*; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D000911:Antibodies, Monoclonal; D006511:Hepatitis B Antigens; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein