A型单端孢霉烯族毒素多克隆抗体及其免疫磁珠制备技术

Polyclonal antibody and immunomagnetic beads preparation technology for type A trichothecenes

目的制备用于隐蔽态A型单端孢霉烯族毒素富集净化的免疫磁珠。方法采用A型单端孢霉烯族(简称A型单族)毒素人工抗原3-乙酰-新茄腐镰刀菌烯醇-酸酐-牛血清蛋白(3-Ac-NEOS-HS-BSA)免疫新西兰长耳白兔,分离血清制备A型单族毒素多克隆抗体,测定效价并纯化。然后优化高效价抗体与免疫磁珠的偶联条件并应用。结果免疫5次后的兔血清中A型单族毒素抗体效价最高为1:64 000。抗体和磁珠的最优偶联条件为:24℃、pH 7.4、0.02 mol/L含10%甲醇的PBS缓冲液中旋转捕获24 h。结论 A型单族毒素多克隆抗体及其免疫磁珠的制备为同步富集净化复杂基质中A型单族毒素、提高方法的检测限奠定基础。

Objective Preparing immunomagnetic beads for enrichment and purification of mask type A trichothecenes.Methods Through immunizing New Zealand white rabbits with family A toxin artificial antigen 3-Ac-NEOS-HS-BSA,the polyclonal antibodies of type A trichothecenes（ type A family） were isolated,as well as its antibody titre were determinated and purified.The coupling condition of high titer antibody and immunomagnetic beads were optimized and applied.Results After 5 times of immunization with type A trichothecene family toxin,the rabbit serum inhibited the highest antibody titer（ 1 ： 64 000）.The optimal coupling condition of antibody and immunomagnetic beads were： temperature = 24 ℃,rotation capture in pH 7.4,0.02 mol/L PBS buffer solution with 10% methanol in 24 h.Conclusion The establishment of preparation method of type A single family toxin polyclonal antibody and immunomagnetic beads can make a standard for enrichment and purification of type A matrix complex single family toxins.