人血清载脂蛋白CⅠ和CⅢ的分离提纯及其单克隆抗体的制备

Isolation and Purification of Human Serum Apolipoprotein CⅠ,CⅢ and Preparation of Their Monoclonal Antibodies

本文应用超速离心、凝胶过滤层析、高效液相色谱法分离纯化了人血清极低密度脂蛋白—载脂蛋白ＣⅠ和ＣⅢ。经等电聚焦电泳、十二烷基硫酸钠—聚丙烯酰胺电泳、双向免疫扩散等方法鉴定了纯度。以载脂蛋白ＣⅠ和ＣⅢ纯品免疫ＢＡＬＢ／Ｃ小鼠，应用淋巴细胞杂交瘤技术建立了４株抗人载脂蛋白ＣⅠ和２株抗人载脂蛋白ＣⅢ杂交瘤细胞。所得单克隆抗体与载脂蛋白ＡⅠ、ＡⅡ、Ｂ、ＣⅡ、Ｅ和白蛋白等无交叉反应。

Apolipoprotein CⅠ,CⅢwere isolated and purified by ultracentrifugation, gel filtration and high performance liquide chromatography from human serum very low density lipoprotein. The purity of apo CⅠ and CⅢ was characterized by IEF、SDS-PAGE and double immunodiffusion. The BALB/C mice were immunized by purified apo CⅠ or CⅢ.4 lines of hybridoma to apo CⅠ and 2 lines of to apo CⅢwhich were able to secreat monoclonal antibOdies(McAb) to human apo CⅠ and apo CⅡ respectively were established. The McAbs have no cross reaction to apo AⅠ,AⅡ,B,CⅡ,E and human albumin.