应用亲和层析纯抗入Apo(a)及ApoBIgG测定人血浆脂蛋白(a)——夹心ELISA法

Enzyme-linked Immunosorbent Assay of Human Plasma Lipoprotein(a)by Using Affinity-purified Anti-human Apo(a)and AopB IgG

利用亲和层析纯抗人 Apo(a) 和 ApoB IgG 在国内首次建立了 Lp(a)测定的夹心 ELISA 方法,此法不受纤溶酶原的干扰,与纤溶酶原、HDL 及 CDL 的交叉反应率均小于0.05%;标准曲线在0.5～0.6 ng/孔范围内呈线性关系,对应于血浆稀释1000倍时 Lp(a)含量5～600 mg/L;最低可测0.3 ng/孔,平均批内和批间变异系数分别为4.2%和7.5%,平均回收为率102%.本法与国外采用Apo(a)单克隆抗体的 Capture ELISA 及 EIA 均有很好的相关性,相关系数 r 分别为0.982(n=50)和0.919(n=60)。用此方法测定208例正常血脂个体.均值为122.17 mg/L (s 为121.63),中位数为90 mg/L.未发现性别差异。

This paper presented a sandwich-type enzyme-linked immunosorbent assay(ELISA)for quantita- tion of human plasma lipoprotein(a)[Lp(a)],which was based on the use of an affinity-purified an- ti-Lp(a)IgG as coating antibody and an anti-ApoB IgG-peroxidase conjugate for detection of bound Lp(a).The ELISA assay,which is insensitive to the presence of plasminogen,detects amounts corre- sponding to Lp(a)contents of 5～600 mg/L in plasma samples diluted 1000-fold.Mean recocery was 102% and mean intra-and inter-assay CVs for assay of 20 samples were 4.2% and 7.5%,respec- tively.Results correlated well with those by Capture ELISA(r=0.982.n=50)and electroim- munoassay(r=0.919,n=60),For 208 normolipidemic subjects,the mean and median Lp(a)con- centration are 122.17 mg/L(s=121.63mg/L)and 90 mg/L respectively.