摘要
目的：制备抗甲状腺素抗体Ｆａｂ’片段，并达到符合酶标记的要求，以期可用于夹心酶联免疫方法（ＥＬＩＳＡ）测定血清中游离甲状腺素的浓度。方法：用硫酸钠ＤＥＡＥ纤维素层析法从兔抗Ｔ４的抗血清中提纯相应的ＩｇＧ，用胃蛋白酶将ＩｇＧ消化为Ｆ（ａｂ’）２片段，后者再用巯基乙醇还原为Ｆａｂ’。结果：用ＳＤＳ聚丙烯酰胺凝胶电泳（ＰＡＧＥ）鉴定纯化的ＩｇＧ、Ｆ（ａｂ’）２、Ｆａｂ’均为单一条带，分子量分别为１５ＫＤ、９７ＫＤ、４９ＫＤ；纯化后得到的ＩｇＧ、Ｆ（ａｂ’）２、Ｆａｂ’的回收率分别为６２．２％、６７．０％、７１．２％；用放免法检测均具有免疫活性；Ｆａｂ’每分子中的巯基数为１．０６。结论：用本文方法制备的Ｆａｂ’片段经鉴定，合乎酶标记和酶免工作的要求。
Objective: To prepare the Fab’ fragment ,used for ELISA from IgG against thyroxine(T 4) Methods: IgG was purified using sodium sulfate saltout/DEAE cellulose chromatography from rabbit anti thyroxine serum The purified IgG was digested with pepsin to produce F(ab’) 2 Then, the F(ab’) 2 was reduced to Fab’ with 2 mercaptoenthanol Results:IgG and their fragment were identified with SDS PAGE and molecular weight(MW) were 14 99KD、97 35KD、48 82KD, respctively The recovery by protein assay were 62 2%、67 0%、71 2%, respectively The number of thiol groups was 1 06 per Fab’ Conclusion: The anti throxine Fab’ fragment prepared can be suitablely used for conjugation with enzyme
出处
《天津医科大学学报》
1998年第4期314-316,共3页
Journal of Tianjin Medical University
基金
天津市教委基金
