title: [Expression of the catalytic antibodies in eukaryotic systems].
abstract: Expression of recombinant antibodies in mammalian cells is one of key problems in immunobiotechnology. Alternatively, expression of a broad panel of antibodies and of their fragments may be effectively done in yeast cells. We obtained expression strains of the methylotrophic beast Pichia pastoris producing single chain human catalytic antibody A17 (A.17scFv), Fab-fragment (A.17Fab) and full-size light chain (A.17Lch). These antibodies were characterized in terms of functional activity. The capacity to specifically bind and transform organophosphorus compounds has been demonstrated for A.17scFv and A.17Fab. The loss of activity of the antibody light chain when expressed alone indicates that the active site is formed by both heavy and light chains of the antibody. We determined the reversible constant Kd and the first order constant (k2) of the reaction of the covalent modification of A.17scFv and A.17Fab by irreversible inhibitor of the serine proteases p-nitrophenyl 8-methyl-8-azobicyclo[3.2.1]phosphonate (Phosphonate X). Calculated values indicate that activity of the antibodies expressed in yeast is similar to the full-size antibody A17 and single chain antibody A.17 expressed in CHO and E. coli cells respectively.
authors.forename: A V
authors.initials: AV
authors.lastname: Zakharov
authors.forename: I V
authors.initials: IV
authors.lastname: Smirnov
authors.forename: M V
authors.initials: MV
authors.lastname: Serebriakova
authors.forename: M A
authors.initials: MA
authors.lastname: Dronina
authors.forename: A V
authors.initials: AV
authors.lastname: Kaznacheeva
authors.forename: I N
authors.initials: IN
authors.lastname: Kurkova
authors.forename: A A
authors.initials: AA
authors.lastname: Belogurov
authors.forename: A
authors.initials: A
authors.lastname: Friboulet
authors.forename: N A
authors.initials: NA
authors.lastname: Ponomarenko
authors.forename: A G
authors.initials: AG
authors.lastname: Gabibov
authors.forename: T V
authors.initials: TV
authors.lastname: Bobik
mesh_terms: D000818:Animals; D017151:Antibodies, Catalytic / Q000096:biosynthesis* / Q000235:genetics; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D006801:Humans; D010843:Pichia; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D017151:Antibodies, Catalytic; D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies