| 抗人层粘连蛋白Fab＇的制备 |
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| 引用本文： | 陈金国,柴明胜,叶仲贤,朱炳钗.抗人层粘连蛋白Fab＇的制备[J].免疫学杂志,1996(1). |
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| 作者姓名： | 陈金国 柴明胜 叶仲贤 朱炳钗 |
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| 作者单位： | 上海海军医学研究所 |
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| 摘 要： | 用ＤＥＡＥ纤维素ＤＥ－５２反相吸附纯化抗人层粘连蛋白（ＬＮ）ＩｇＧ，ＩｇＧ回收率达１１．６ｍｇ／ｍｌ血清。胃蛋白酶消化ＩｇＧ成Ｆ（ａｂ＇）２，再用１０ｍｍｏｌ／Ｌ２－巯基乙醇３７℃反应９０ｍｉｎ还原成Ｆａｂ＇，回收率约８１％，放免效价为１：３５００．测得纯化的ＩｇＧ、Ｆ（ａｂ＇）２及Ｆａｂ＇分子量分别为１５００００、９００００、４５０００，与理论值相近。放免检测中用Ｆａｂ＇取代抗血清可纠正对血清ＬＮ测定的误差，从而建立检测ＬＮ的准确、灵敏方法。
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| 关 键 词： | Fab＇片段，单价抗体，层粘连蛋白，放射免疫分析 |
Preparation of Fab' fragment from rabbit anti-human laminin serum |
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| Chem Jinguo,Chai Mingsheng,Ye Zhongxian,Zhu Binchai.Preparation of Fab' fragment from rabbit anti-human laminin serum[J].Immunological Journal,1996(1). |
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| Authors: | Chem Jinguo Chai Mingsheng Ye Zhongxian Zhu Binchai |
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| Abstract: | About 11.6mg IgG was purified from 1 ml rabbit anti-human laminin serum using DE-52 cellulose reverse absorbent chromatography.The concentrated IgG was digested with pepsin to produce F(ab')2,then the F(ab') 2 was reduced to Fab'with 10 mmol/L 2-mercaptoenthanol for 90min at 37.The recovery of Faywas 81%,and the titer by RIA was 1:3500.The MWs of purified IgG,F(ab') 2 and Fab' were 150000,90000 and 45 000,respectively.The use of Fab' in RIA instead of the whole antiserum could obviate the nonparallel inhibition profiles for serum samples and standard laminin so that a reliable assay for the laminin in serum can be developed. |
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| Keywords: | Fab' fragment Monovalent antibody Laminin Radioimmunoassay |
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