人血液血管细胞生成素的胎肝免疫印记检测  

IMMUNO-BLOT DETECTION OF HEMANGIOPOIETIN IN THE HUMAN FETAL LIVER

作　　者：秦迎松[1] 韩忠朝[2] 

机构地区：[1]徐州医学院病理生理学教研室,江苏徐州221002 [2]中国医学科学院血液学研究所泰达生命科学技术研究中心,天津300457

出　　处：《中国应用生理学杂志》2007年第1期102-105,共4页Chinese Journal of Applied Physiology

基　　金：国家863项目资助课题(2001AA215311;2002AA223354);国家973项目资助(001CB5101);国家科技部95重大资助项目(95-special-10)

摘　　要：目的:制备一种新蛋白——人血液血管细胞生成素(hemangiopoietin,HAPO)的单克隆抗体,检测其在胎肝中的表达。方法:杂交瘤方法制备抗HAPO的单克隆抗体;非竞争酶联免疫吸附实验测定抗体的相对亲和力;蛋白G亲和层析纯化腹水中的抗体,免疫印记方法检测胎肝中天然HAPO的表达。结果:所获单抗分别为IgG1及IgM,其轻链均为κ。三株IgG1亚类单抗的相对亲和力分别为3.06×109mol/L,6.07×108mol/L和1.71×1010mol/L。亲和纯化后抗体的纯度达99%以上。胎肝中在蛋白水平上可以检测到HAPO的表达,天然HAPO的分子量接近于重组HAPO的分子量。结论:人胚胎肝组织中在蛋白水平上可以检测到HAPO的表达。Aim. To detect the expression of a novd protein, hemangiopoietin （HAPO）, in the human fetal liver at the protein level. Methods. Monoclonal antibodies （moAbs） against HAPO were produced by traditional hybridoma technique. Their affinities were calculated from the results of non-competitive ELISA and the antibodies were purified by protein G affinity chromatography. Expression of in the liver was detected by SDS-PAGE and Western blot. Results： Five strains of moAbs were screened out in total, among which three were IgG1 and the other two were IgM. Their light trains were all belonged to κ. The rdative affinities of the three IgG1 form moAbs were 3.06 ×10^9 mol/L, 6.07 ×10^8 mol/L and 1.71 ×10^10 mol/L respectively. After purification, the purity of the moabs could reach more than 99 %. HAPO expression was detected at the protein level in the human fetal liver, and the apparent molecular weight of the nature HAPO was very close to but a little higher than our recombinant one. Conclusion： HAPO was expressed in the human fetal liver at the protein level.

关 键 词：人血液血管细胞生成素 单克隆抗体 免疫印记 

分 类 号：Q781[生物学—分子生物学]