摘要
研制一种蛋白 C(PC)的特异单克隆抗体(McAb),它可以识别在钙离子存在与否的条件下呈现不同构象的 PC 抗原,将其偶联至 Sepharose 4B 凝胶上,组成亲和层析柱。凝胶上的 McAb 能连接无钙离子血浆中的 PC,这种免疫纯化法一步即可达到12000倍的纯化。PC 成品的比活性为180～230U/mg,在非还原的 SDS-PAGE 图谱上呈现两条带(α-PC 和β-PC),在还原的 SDS-PAGE 图谱上呈现5条带(α-SC-PC,β-SC-PC,α-HC,β-HC 和 LC)。
A special monoclonal antibody(McAb)to protein C(PC)was prepared,itrevealed the different conformation in the absence and presence of Ca^(++).This McAbwas coupled to Sepharose-4B and the gel was loaded into the column for thepurification of PC.The immobilized antibody was capable of binding PC fromthe plasma in the absence of Ca^(++).This immunopurification resulted in a 12 000-fold after one step proceduce.The specific activity of the PC product ranged be-tween 180-230 U/mg and behaved as two bands(α-PC andβ-PC)on nonreducedSDS-PAGE and five bands(α-S-PC,β-S-PC,α-heavy chain,β-heavy chainand light chain)on reduced SDS-PAGE.
出处
《中国输血杂志》
CAS
CSCD
1993年第1期1-5,共5页
Chinese Journal of Blood Transfusion
关键词
蛋白C
单克隆抗体
免疫亲和层析
Protein C
Monoclonal Anibody
Immunoaffinity chromatography
Calcium Ion
