rac-Tetra(m-nitrobenzene)porphyrin, H2TNO2PP, synthesized from o-nitro-benzaldehyde and pyrrole was reduced to H2TamPP, from which tetra(o-phenylacetyl benzene) porphyrin and especially its α,α,α,α-isomer were obtained. The α,α,α,α-isomer was used as hapten to immunize BALB/C mice. The latter gave antibody value of 320 as determined by ELISA method, indicating that the artificial hapten could induce the production of antibody without use of carrier.
The meso-tetra(α,α,α,α-O-phenylacetylbenzene)porphyrin was used as a complete antigen to elicit monoclonal antibody 1F2 through the immunization and cell fusion techniques. The antibody could combine with Mn porphyrin and showed a haloperoxidase activity. The affinities of the antibody for antigen and Mn porphyrin were 4 98×10 -5 mol/L and 7 63×10 -5 mol/L, respectively. When the antibody-Mn porphyrin complex was used to catalyze the halogenation reaction, the Michaelis constant and reactive rate constant κ cat for H 2O 2 were 5 127 0×10 -3 mol/L and 0 483 4 s -1, respectively. The enzyme exchange number κ cat/K m=94 285 1 mol/(L·s).