抗胸腺素α1(Tα1)多克隆抗体的纯化及转基因藻中Tα1的检测  

Purification of Polyclonal Antibody to Thymosin α1and Detection of Tα1 in Transgenic Algae

作　　者：黄小花[1] 周克夫[1] 章军[1] 刘仁海[1] 徐虹[1] 

机构地区：[1]厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2004年第B08期122-125,共4页Journal of Xiamen University：Natural Science

基　　金：国家海洋863课题(819 04 03)资助

摘　　要：采用不同浓度的硫酸铵分级盐析纯化抗Tα1多克隆抗体,然后用不同量的牛血清白蛋白(BSA)与兔抗Tα1抗体进行中和,以去除血清中的抗BSA抗体,并利用纯化后的抗Tα1多克隆抗体检测转基因藻中Tα1.结果表明:采用25%(NH4)2SO4进行分离纯化,并将BSA加入到血清中进行中和,37℃处理30min,BSA的量与血清的比例以1μL血清+15μgBSA可达到中和血清中的抗BSA抗体的目的.获得的抗Tα1抗体效价与未进行分离前基本相同,并且阴性本底低.采用ELISA方法检测野生藻和转基因藻中的Tα1,转基因藻的Tα1OD450值比野生藻的高2.1倍以上,提示Tα1在转基因藻中成功表达.实验证明,此方法能有效纯化抗胸腺素α1多克隆抗体,获得特异性强的抗胸腺素Tα1抗体,并且效价不受影响.We separated and purified polyclonal antibody to thymosin α1 using different concentrations of(NH_4)_2SO_4,then neutralized Tα1 antibody of rabbits so as to wipe off BSA antibody in serum using different quantities of BSA,and detected Tα1 in transgenic algae with purifies polyclonal antibody to thymosin α1.The result proved that the best conditions were:with 25%(NH_4)_2SO_4 to salt out,neutralization with BSA at 37℃ for 30 min,the satisfaction ratio between serum and BSA was 1 μL serum,15 μg BSA at 37℃ for 30 min,and we got a satisfactied result with this antibody to detect the Tα 1 in transgenic algae.The detection showed that the OD_(450) value of transgenic alga was over two times as that of wild type algae,and indicated that Tα1 was expressed in transgenic algae successfully.The purified antibody had properties of strong specification,high sensitivity,and steady antibody titeration.

关 键 词：抗胸腺素α1 酶联免疫吸附反应 分离 纯化 抗体 转基因藻 

分 类 号：R392.11[医药卫生—免疫学]