title: Antiperoxidase antibodies enhance refolding of horseradish peroxidase.
abstract: The effect of monoclonal antibodies on protein folding was studied using horseradish peroxidase refolding from guanidine hydrochloride as a model process. Among the five antiperoxidase clones tested, one was found to increase the yield of catalytically active peroxidase after guanidine treatment. The same clone also increased the activity of the native peroxidase by a factor of 2-2.5. While peroxidase refolding under standard conditions resulted in the recovery of only 7-8% of the initial catalytic activity, antibody-assisted refolding increased the yield to 50-100% (or 20-40% from the activity of native enzyme with antibodies). Kinetics of autorefolding and antibody-assisted refolding differed significantly. In the course of autorefolding the catalytic activity was recovered within the first 2.5 min and did not change further within a 2.5- to 60-min interval, whereas in the course of antibody-assisted refolding maximal catalytic activity was attained only in 60 min. The yield of active peroxidase for the antibody-assisted refolding depended linearly on the antibody concentration. The observed effect was strongly specific. Other antiperoxidase clones tested as well as nonspecific antithyroglobulin antibody affected neither kinetics, no the yield of peroxidase refolding.
authors.affiliation: Institute of Biochemistry, Russian Academy of Sciences, Leninsky Prospect 33, 117071 Moscow, Russia.
authors.forename: D N
authors.initials: DN
authors.lastname: Ermolenko
authors.forename: A V
authors.initials: AV
authors.lastname: Zherdev
authors.forename: B B
authors.initials: BB
authors.lastname: Dzantiev
authors.forename: V O
authors.initials: VO
authors.lastname: Popov
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000918:Antibody Specificity; D019791:Guanidine / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D007700:Kinetics; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding
chemical_list: D000911:Antibodies, Monoclonal; D006735:Horseradish Peroxidase; D019791:Guanidine