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Preparation and Characterization of a Chloroperoxidase-like Catalytic Antibody
College of Life Science and Agriculture, Key Laboratory of Tropic Biological Resources of Ministry of Education, Hainan University, Haikou 570228, P. R. China
College of Chemistry, Huazhong Normal University, Wuhan 430079, P. R. China
Research Center for Wildlife Animal Diseases, Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, P. R. China
College of Life Science, Huazhong Normal University, Wuhan 430079, P. R. China
* Authors to whom correspondence should be addressed.
Received: 10 February 2007; in revised form: 12 April 2007 / Accepted: 16 May 2006 / Published: 29 May 2007
Abstract: The small molecule, meso-tetra(α,α,α,α-o-phenylacetamidophenyl)porphyrin(Mr1147.0) was used as complete antigen to elicit MAb through the immunization and cellfusion techniques.The MAb 1F2 obtained was demonstrated to be very pure byMALDI/TOFMS.The subtype of MAb 1F2 is IgG2a, which has a relative molecular weightof 156,678.8 Da.No significant change in the intensity of absorption peaks in UV and CDspectra was observed over a pH range between 6 and 12.The high stability of the abzymeand the tight binding between Fe porphyrin and antibody were also demonstrated.Vmax, Km,κcat, κcat/Km for abzyme are 5.18 × 10-8 Ms-1, 1.50 × 10-8M , 0.518 s-1, 3.45 × 107 M-1s-1, respectively.The data obtained indicate that catalytic antibody has highcatalytic activity.The chloroperoxidase activity of MAb 1F2-Fe porphyrin complex is stablefrom 10 °C to 60 °C.
Keywords: meso-Tetra(α,α,α,α-o-phenylacetamidophenyl)porphyrin; MAb; Complete antigen; Chloroperoxidase; Abzyme
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Wang, F.; Huang, X.; Du, L.; Li, W.; He, H.; Qi, C. Preparation and Characterization of a Chloroperoxidase-like Catalytic Antibody. Int. J. Mol. Sci. 2007, 8, 445-454.
Wang F, Huang X, Du L, Li W, He H, Qi C. Preparation and Characterization of a Chloroperoxidase-like Catalytic Antibody. International Journal of Molecular Sciences. 2007; 8(5):445-454.
Wang, Fengyang; Huang, Xueying; Du, Li; Li, Weiguo; He, Hongxuan; Qi, Chao. 2007. "Preparation and Characterization of a Chloroperoxidase-like Catalytic Antibody." Int. J. Mol. Sci. 8, no. 5: 445-454.