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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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