石房蛤毒素免疫亲和柱制备及柱效测试研究
目的 阐明石房蛤毒素(saxitoxin, STX)免疫亲和柱对11种麻痹性贝类毒素的亲和作用。方法 通过纯化的STX单克隆抗体与琼脂糖凝胶(sepharose 4B)制备STX免疫亲和柱, 采用11种麻痹性贝类毒素(paralytic shellfish poisoning, PSP)标液进行过柱实验, 优化上柱条件, 采用液相色谱-质谱串联法进行毒素检测。结果 STX免疫亲和柱对新石房蛤毒素(neosaxitoxin, neo-STX)、脱氨甲酰基新石房蛤毒素(decarbamoylneosaxitoxin dihydrochloride, dcneo-STX)、膝沟藻毒素1(gonyautoxin-1, GTX1)、膝沟藻毒素4(gonyautoxin-4, GTX4)基本没有亲和力作用; 而对7种PSP的亲和力强弱顺序为: STX>N-磺酰氨甲酰基类毒素5(gonyautoxin-5, GTX5)>脱氨甲酰基石房蛤毒素(decarbamoylsaxitoxin dihydrochloride, dcSTX)>膝沟藻毒素3(gonyautoxin-3, GTX3)>脱氨甲酰基膝沟藻毒素3(decarbamoylgonyautoxin-3, dcGTX3)>脱氨甲酰基膝沟藻毒素2(decarbamoylgonyautoxin-2, dcGTX2)>膝沟藻毒素2(gonyautoxin-2, GTX2), 其中对STX、GTX5、dcSTX、GTX3的回收率为61.2%~99.0%。结论 该STX免疫亲和柱对STX、GTX5、dcSTX、GTX3有较好的吸附效果, 能够满足样品检测前处理的要求, 为水产品中麻痹性贝类毒素的提取方法研究提供了新技术的参考依据
Objective To clarify affinity effect of saxitoxin (STX) immunoaffinity column (IAC) on 11 species paralytic shellfish poisoning (PSP). Methods STX immunoaffinity column was prepared by purified STX monoclonal antibody and sepharose 4B. Eleven kinds of paralytic shellfish poisoning (PSP) standard liquid were used for column experiment, the conditions of upper column were optimized, and PSP was detected by liquid chromatography-mass spectrometry. Results STX IAC had no affinity effect on neosaxitoxin (neo-STX), decarbamoylneosaxitoxin dihydrochloride (dcneo-STX), gonyautoxin-1 (GTX1), gonyautoxin-4 (GTX4) basically, however it had relative strong effect on the other 7 kinds of PSP and the affinity order was: STX>GTX5>dcSTX>GTX3>dcGTX3>dcGTX2>GTX2. The recoveries of STX, GTX5, dcSTX, and GTX3 were 61.2%?99.0%. Conclusion The STX IAC has good adsorption effect on STX, GTX5, dcSTX, and GTX3, which can satisfy the demands of sample pre-treatment, and provide new technology reference and support for the PSP extraction research in aquatic products.
标题：石房蛤毒素免疫亲和柱制备及柱效测试研究
英文标题：Preparation and column efficiency research of saxitoxin immunoaffinity column
作者：
岳亚军 深圳市罗湖区疾病预防控制中心
朱波 深圳市罗湖区疾病预防控制中心
伦丽丽 北京美正生物技术有限公司
徐娜 北京美正生物技术有限公司
梁玉波 国家海洋环境监测中心
中文关键词:石房蛤毒素,麻痹性贝类毒素,免疫亲和柱,高效液相色谱-串联质谱法,回收率,
英文关键词:saxitoxin,paralytic shellfish poisoning,immunoaffinity column,high performance liquid chromatography- tandem mass spectrometry,recovery,
发表日期：2019-04-08
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