PMID: 9647821Jul 2, 1998Paper

Development of a streptavidin-conjugated single-chain antibody that binds Bacillus cereus spores

Applied and Environmental Microbiology
K KooH E Swaisgood

Abstract

Control of microorganisms such as Bacillus cereus spores is critical to ensure the safety and a long shelf life of foods. A bifunctional single chain antibody has been developed for detection and binding of B. cereus T spores. The genes that encode B. cereus T spore single-chain antibody and streptavidin were connected for use in immunoassays and immobilization of the recombinant antibodies. A truncated streptavidin, which is smaller than but has biotin binding ability similar to that of streptavidin, was used as the affinity domain because of its high and specific affinity with biotin. The fusion protein gene was expressed in Escherichia coli BL21 (DE3) with the T7 RNA polymerase-T7 promoter expression system. Immunoblotting revealed an antigen specificity similar to that of its parent native monoclonal antibody. The single-chain antibody-streptavidin fusion protein can be used in an immunoassay of B. cereus spores by applying a biotinylated enzyme detection system. The recombinant antibodies were immobilized on biotinylated magnetic beads by taking advantage of the strong biotin-streptavidin affinity. Various liquids were artificially contaminated with 5 x 10(4) B. cereus spores per ml. Greater than 90% of the B. cereus spore...Continue Reading

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Citations

Dec 25, 2008·Analytical and Bioanalytical Chemistry·Tae Jung ParkSang Yup Lee
Apr 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Bin ZhouKim D Janda
Sep 28, 2001·Biochemical and Biophysical Research Communications·J G BrunoC A Batt

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