Preparation and Comparative Analysis of β-glucosidase Antibodies
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摘要: β-葡萄糖苷酶是一种广泛存在于自然界的水解酶,其含量高低直接关系到水解产物的量。木薯含有大量的生氰糖苷,是β-葡萄糖苷酶的底物之一,被水解后将产生有毒的氰化物,给食品安全带来隐患。为保障食品安全,给木薯及其制品的氰化物含量提供快速检测技术。本研究以A,B,C 3种来源的β-葡萄糖苷酶为抗原免疫Balb/C小鼠,并制备单克隆抗体和做出初步评价。结果表明,A抗原免疫的小鼠融合后获得5株杂交瘤细胞株,效价均达到105以上,抗体亚型均为IgG1型,但单克隆抗体与B抗原、C抗原、KLH、BSA均无交叉反应。说明不同来源的β-葡萄糖苷酶免疫小鼠得到的抗体均为特异性抗体。Abstract: β-glucosidase is a hydrolase which exists widely in nature, and its content is directly related to the amount of hydrolysate. Cassava contains a large number of cyanogenic glycosides which are one of the substrates of β-glucosidase and will produce a poisonous substance, cyanide after hydrolyzed by β-glucosidase, bringing hidden dangers to the food safety. For food safety, an attempt was made to provide a rapid detection technique for cyanide content of cassava and its products. The Balb/C mice were immunized with β-glucosidase from A, B and C sources as antigens, and their monoclonal antibodies were prepared and evaluated. The results showed that the antibodies obtained from the mice immunized with the three sources（A, B and C） of β-glucosidase did not cross-react with each other. Five hybridoma cell lines were obtained from the spleen of the mice immunized with Antigen A, and the titer of antibodies was above 105. All the subtypes were IgG1, but all the monoclonal antibodies had no cross-reaction with the Antigens B and C, KLH and BSA. It was concluded that the antibodies obtained from the mice immunized with different sources of β-glucosidase were specific.
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Key words:
- β-glucosidase /
- antibody /
- antibody preparation
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