Development and Application of a Piezoimmunosensor for Staphylococcal Enterotoxins

• Main Authors: LIN, YI-CHING, 林依晴
• Other Authors: Chen Chein-Yuan
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/59817802623716570405

碩士 === 國立臺灣大學 === 農業化學系研究所 === 86 === In this study, SEA (staphylococcal enterotoin A), SEB, SEC3 were purified from the cultural broth of Staphylococcus aureus CCRC 13966, 13825, 13828, respecti vely. The anti-SEs antibodies were obtained from ascites of mice andfurther pu rified by Protein A affinity column chromatography. The measurementof piezoele ctric sensor was set up with a university counter and a 10 MHzquartz crystal o scillated by a SN 7400-TTL. The crystal was dipped in a 2 % o-Phenylenediamine solution for 15 sec, then immersed in a 2.5 % Glutaraldehydesolution for 30 m in, and then immersed in a solution of 50 ug/ml antibody in10 mM phosphate buf fer (pH 7.0) at 40 C for 1 hr. The crystal surface couldbe coated with optimum amount of antibody. A 5 to 10 min incubation time wasenough for the immobiliz ed antibody to take up antigen. Using this piezo-immunosensor, the frequency s hift was linearly correlated with SEA or SEB inthe range of 5 pg/ml to 5 ng/ml , and with SEC3 in 5 pg/ml to 50ng/ml.The average recovery of SEs added to unc ontaminated milk was 60 %. Repeated usageof the antibody coated crystal could be obtained by pH 2.5, 0.5 M or pH 3.0, 0.1 M Glycine-HCl buffer.