title: Purification and characterization of mouse single-chain antibody against polycyclic aromatic hydrocarbons.
abstract: Polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyren mainly induce lung cancer in humans. We characterized the mouse single chain antibody against benzo[a]pyren (pSh). pSh was expressed and purified as cellulose binding domain fusion (pSh-CBD). The pSh-CBD bound five different PAH with high affinity. The 18 amino acid linker connected pSh-CBD heavy and light chains provided correct protein folding. The KDs for pSh-CBD and polycyclic aromatic hydrocarbons were similar to KDs for monoclonal antibody, approximately 10(-8). Separately heavy and light chains of pSh-CBD did not interact with benzo[a]pyren. Previously defined eleven pSh-CBD aa involved to benzo[a]pyren binding were confirmed by mutagenesis.
authors.affiliation: a Laboratory of Biotechnology , The Institute of Human Ecology of Siberian Branch of Russian Academy of Science , Kemerovo , Russia.
authors.forename: Valentin A
authors.initials: VA
authors.lastname: Ustinov
authors.forename: Anton V
authors.initials: AV
authors.lastname: Averjanov
authors.forename: Andrey N
authors.initials: AN
authors.lastname: Glushkov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D001564:Benzo(a)pyrene / Q000737:chemistry* / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D057127:Single-Chain Antibodies; D001564:Benzo(a)pyrene; D002482:Cellulose
keywords: antibody; benzo[a]pyrene; immunology; polycyclic aromatic hydrocarbons; proteins; single-chain antibody