鸡血浆纤维连接蛋白的提纯及酶联免疫吸附检测方法的建立

PURIFICATION OF CHICKEN PLASMA FIBRONECTIN AND ENZYMELINKED IMMUNOSORBENT ASSAY TO QUANTIFY FIBRONECTIN

利用纤维连接蛋白 (FN)与明胶特异结合的特点 ,用明胶亲和层析结合电泳裁胶的方法提纯鸡血浆FN并制备了兔抗鸡FN抗血清。纯化的FN经凝胶电泳鉴定为一条蛋白带且与人FN在同一位置上 ,亚单位分子量约 2 3 0KDa ;经免疫鉴定 ,提纯鸡血浆FN与兔抗人FN抗体有交叉反应性。用兔抗鸡FN抗体包被捕捉抗原 ,建立了检测鸡血浆FN的双抗体酶联免疫吸附法(ELISA) ,此法具有操作简单、特异性强、灵敏度高的特点 ,适用于鸡血浆FN的检测。鸡血浆FN的提纯及其检测方法的建立 ,为FN在兽医领域的进一步研究和应用奠定了基础。

WT5BZ]Through the use of fibronectin′s property of binding to gelatin,chicken plasma fibronectin (FN) was purified by improved gelatin-affinity chromatography and preparative gel electrophoresis.Rabbit anti-chicken plasma FN was made.Purified FN was identified to be a single major band with a molecular weight of about 230 KDa and be on the same position of human FN by SDS-polyacrylamide gel electrophoresis.FN kept primary antigen property and could cross-react with rabbit anti-human FN antibody.Wells were coated with antibody,and an double antibodies enzyme-linked immunosorbent assay method was established to quantify chicken plasma FN.The method was characterized by being easy to operate,strong specificness,high sensitivity and was suitable to assay soluble FN.Purification and establishment of assay method of chicken plasma FN will do good to further research and use in the filed of veterinary medicine. [WT5HZ]