Thermostable peroxidase activity with a recombinant antibody L chain‐porphyrin Fe(III) complex
- 20 November 1995
- journal article
- Published by Wiley in FEBS Letters
- Vol. 375 (3) , 273-276
- https://doi.org/10.1016/0014-5793(95)01224-3
Abstract
In order to engineer a new type of catalytic antibody, we attempt to use a monoclonal antibody L chain as a host protein for a porphyrin. TCPP (meso‐tetrakis(4‐carboxyphenyl)porphyine) was chemically synthesized and Balb/c mice were immunized using TCPP as a hapten. Two hybridoma cells (03‐1, 13‐1), that produce monoclonal antibody against TCPP, were obtained. Genes for both H and L chains of monoclonal antibodies were cloned, sequenced and overexpressed using E. coli as a host. ELISA and fluorescence quenching method show that the independent antibody L chains from both Mab03‐1 and Mab13‐1 have specific interaction with TCPP. Furthermore, the recombinant antibody L chain from Mab13‐1 exhibits much higher peroxidase activity than TCPP Fe(III) alone. The enzyme activity was detectable with pyrogallol and ABTS (2,2‐azinobis‐3‐ethylbenzthiazolin‐6‐sulfonic acid) but not with catechol. This new catalytic antibody was extremely thermostable. Optimum temperature of the peroxidase reaction by the complex of 13‐1L chain and TCPP Fe(III) was 90°C, while that the TCPP Fe(III) alone was 60°C.Keywords
This publication has 16 references indexed in Scilit:
- Cooperative Binding of Porphyrin by Anti-Porphyrin AntibodiesChemistry Letters, 1991
- Peroxidase activity of an antibody-heme complexJournal of the American Chemical Society, 1990
- An antibody-catalyzed bimolecular Diels-Alder reactionJournal of the American Chemical Society, 1990
- Antibody-catalyzed redox reactionJournal of the American Chemical Society, 1989
- Metalloselective anti-porphyrin monoclonal antibodiesJournal of the American Chemical Society, 1989
- A new strategy for the generation of catalytic antibodiesNature, 1989
- Photosensitized cleavage of a thymine dimer by an antibodyJournal of the American Chemical Society, 1988
- Catalytic antibodiesJournal of the American Chemical Society, 1987
- Selective Chemical Catalysis by an AntibodyScience, 1986
- Catalytic AntibodiesScience, 1986