脂质体免疫裂解试验检测人血清胰岛素的初步研究

Homogeneous determination of insulin in human serum using liposome immunolysis assay(LILA):A preliminary study

建立补体介导的脂质体免疫裂解试验（ＬＩＬＡ）以检验人血清中胰岛素含量。应用逆相蒸发法（ＲＥＶ）制备包裹有丽丝胺罗丹明Ｂ的大单层脂质体（ＬＵＶｓ），通过碳二亚胺（ＥＤＣＩ）法偶联胰岛素抗原，当加入胰岛素抗原、一定量抗体，在补体参与下，建立竞争抑制测定模式。结果，丽丝胺罗丹明Ｂ吸光度值的改变与血清中胰岛素浓度呈线性关系，回归方程为，Ｙ＝２２７．７２－８０．４４１Ｘ（ｒ＝－０．９９６６），批内变异系数（ＣＶ）为３．８％～８．５％，批间ＣＶ９．３％～１３．０％，回收率为８３．４％～１１４．０％，灵敏度为１．２５μＵ／ｍｌ，ＬＩＬＡ（Ｙ）和ＲＩＡ（Ｘ）相关显著，Ｙ＝１．０３Ｘ＋０．４３（ｒ＝０．９７７，Ｐ＜０．００１）。

To develope complement mediated liposome immunolysis assay(LILA)for detecting insulin concentration in human serum.Reverse phase evaporation (REV)method to prepare large unilamellar vesicles(LUVs)encapsulated sulforhodamine B,and obtained immunoliposome by using 1 ethy1 3(3 dimethylaminopropy1)carbodiimide(EDCI)which made insulin couple the LUVs.Competing inhibition assay could be achieved upon the addition of free insulin and anti insulin antibody in the presence of complement.The results revealed that there was a excellent negative correlation between the absorption change of sulforhodamine B and insulin concentration in serum( r =-0 9966). The within run CV and between run CV were 3 8%￣8 5%( n =12),9 3%￣13 0%( n =6)respectively.The recovery was 83 4%￣114 0%.the sensitivity was 1 25 μU/ml. The correlation between LILA( Y ) and RIA (X )was significant: Y=1 03X+0 43(r=0 977,P <0 001).LILA is simple, rapid, specific,sensitive and stable method which is worthy to be studied and popularized.