Article

Acta Biochim Biophys Sin 2005,37:447-452

doi:10.1111/j.1745-7270.2005.00064.x

Preparation and Preliminary Application of Monoclonal Antibodies against Trichokirin-S1, a Small Ribosome-inactivating Peptide from the Seeds of Trichosanthes kirilowii

Xin-Xiu YANG, Feng LI, Wei-Guo HU, Heng-Chuan XIA, and Zu-Chuan ZHANG*

Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences Chinese Academy of Sciences, Shanghai 200031, China

Abstract Trichokirin-S1, a small ribosome-inactivating peptide recently purified from the seeds of Trichosanthes kirilowii, has potential clinical applications because of its small molecular mass. Two stable strains of hybridomas (1F11 and 2A5) that can secrete highly specific monoclonal antibodies (mAbs) against Trichokirin-S1 have been developed using the hybridoma technique. The isotypes of these two mAbs, 1F11 and 2A5, were determined to be IgG2a and IgG1, respectively. The affinity constants, which were measured by non-competitive ELISA, were found to be 2.3×108 M–1 and 2.8×108 M–1, respectively. An immunoaffinity method using 2A5-coupled Sepharose 4B was successfully developed to purify Trichokirin-S1. These two antibodies have also been used to detect Trichokirin-S1 in Western blot.

Key words Trichokirin-S1; monoclonal antibody; Western blot; affinity chromatography

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