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巴沙鱼、博氏巨鲶和低眼无齿𩷶特异性单克隆抗体的制备与鉴定

Production and characterization of monoclonal antibodies specific to pangasius catfish, basa, and tra.

作者信息

Gajewski K G, Chen Y-T, Hsieh Y-H P

机构信息

Department of Nutrition, Food and Exercise Sciences, Florida State Univ., Tallahassee, FL 32306-1493, USA.

出版信息

J Food Sci. 2009 Apr;74(3):C241-7. doi: 10.1111/j.1750-3841.2009.01097.x.

Abstract

Four IgG (subclass IgG1) class monoclonal antibodies (MAbs) strongly reactive to Asian farm-raised Pangasius catfish, tra (Pangasius hypophthalmus) and basa (Pangasius bocourti), have been developed. These MAbs were raised by immunizing an animal with thermal-stable crude sarcoplasmic protein extract of cooked tra. The MAbs were selected by screening hybridoma clones against more than 70 common fish and meat protein extracts. Two MAbs, T7E10 and T1G11, were found to be specific to the Asian Pangasius catfish, tra, and basa, with no cross-reactions with any of the common fish and meat species or with the food additive proteins (bovine serum albumin, soy proteins, milk proteins, egg proteins, and gelatin) tested. MAb T7E10 recognized 2 antigenic proteins (molecular weight approximately 36 and 75 kDa) in raw and cooked tra and basa extracts, while T1G11 bound to several proteins (molecular weight between 13 and 18 kDa) in tra and basa extracts. Two other MAbs, F7B8 and F1G11, recognized a common protein (36 KDa) and cross-reacted with all the fish extracts tested and with several mammalian species. These MAbs can be employed individually or in combination in various formats of immunoassays for rapid identification of Pangasius catfish, either raw or cooked. They can also be used to study the biological, biochemical, and physiological aspects of thermal-stable antigenic proteins. This is the first study identifying these thermal-stable antigenic proteins present in Pangasius catfish as species-specific biomarkers.

摘要

已研发出四种对亚洲养殖的巴沙鱼、苏氏圆腹芒鲶(Pangasius hypophthalmus)和博氏巨鲶(Pangasius bocourti)具有强烈反应性的IgG(亚类IgG1)类单克隆抗体(MAb)。这些单克隆抗体是通过用煮熟的苏氏圆腹芒鲶的热稳定粗肌浆蛋白提取物免疫动物而产生的。通过针对70多种常见鱼类和肉类蛋白提取物筛选杂交瘤克隆来选择这些单克隆抗体。发现两种单克隆抗体T7E10和T1G11对亚洲巴沙鱼、苏氏圆腹芒鲶和博氏巨鲶具有特异性,与任何常见鱼类和肉类物种或所测试的食品添加剂蛋白(牛血清白蛋白、大豆蛋白、牛奶蛋白、鸡蛋蛋白和明胶)均无交叉反应。单克隆抗体T7E10在生的和煮熟的苏氏圆腹芒鲶和博氏巨鲶提取物中识别出2种抗原蛋白(分子量约为36和75 kDa),而T1G11与苏氏圆腹芒鲶和博氏巨鲶提取物中的几种蛋白(分子量在13至18 kDa之间)结合。另外两种单克隆抗体F7B8和F1G11识别一种共同蛋白(36 kDa),并与所有测试的鱼类提取物以及几种哺乳动物物种发生交叉反应。这些单克隆抗体可单独或组合用于各种免疫测定形式,以快速鉴定生的或煮熟的巴沙鱼。它们还可用于研究热稳定抗原蛋白的生物学、生物化学和生理学方面。这是首次将巴沙鱼中存在的这些热稳定抗原蛋白鉴定为物种特异性生物标志物的研究。

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