title: High-level production of a single chain antibody against anthrax toxin in Escherichia coli by high cell density cultivation.
abstract: Previously, we isolated the M18 scFv, which is an affinity matured antibody against the anthrax toxin PA, and observed that its single chain antibody (scAb) form (M18 scAb) exhibited superior stability compared to the scFv. Here, we report high cell density cultivations for preparative scale production of M18 scAb in a 3.5 L fermenter. Briefly, a pH-stat feeding strategy was employed in fed-batch cultivation, and four different cell densities (OD600 of 40, 80, 120, and 150) were examined for the induction of scAb gene expression. Among the four cell densities investigated, lower cell densities (OD600 of 40) showed higher post-induction cell growth and production yields (665 mg/L of scAb). Even though lower solubility (51%) of scAb was achieved at lower cell density (OD600 of 40), monomeric scAb could be purified with high purity (> 95%) using simple purification procedures. The purified scAb from high cell density cultures showed biological activity equivalent to that of scAb purified from shake flask cultivation.
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, Daejeon, 305-701, Republic of Korea. kjjeong@kaist.ac.kr
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
authors.forename: Mridula
authors.initials: M
authors.lastname: Rani
mesh_terms: D000942:Antigens, Bacterial / Q000276:immunology*; D001427:Bacterial Toxins / Q000276:immunology*; D001431:Bacteriological Techniques / Q000379:methods*; D002452:Cell Count / Q000379:methods; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D003470:Culture Media; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C030325:anthrax toxin