反式脂肪酸中反式油酸多克隆抗体的制备

Preparation of Polyclonal Antibody Against Elaidic Acid in Trans Fatty Acids

目的:制备具有特异性的反油酸多克隆抗体。方法:用反式脂肪酸中的反式油酸偶联牛血清白蛋白合成抗原,用一定剂量的抗原对小鼠进行腹腔注射免疫,获得特异性反式油酸多克隆抗体。采用红外光谱检测抗原的合成效果,间接ELISA和蛋白质电泳等方法检验多克隆抗体的效价。结果:使用间接ELISA法和竞争性ELISA法检测获得第4次免疫的抗体,其效价8 100,而血清竞争性可达29.20%。结论:本实验中获得的抗体血清效价不是很高,但竞争性较高,说明血清中存在反式油酸的多克隆抗体。本结果为分离纯化反式脂肪防酸抗体,建立酶联免疫吸附法检测食品中反式脂肪酸,奠定理论依据。

Objective：To prepare the specific anti-elaidic acid polyclonal antibody.Method：Polyclonal antibodies against elaidic acid in trans fatty acids were produced from rats,which were injected by intra-peritoneal injection with a rule of dose and time of elaidic acid conjugated with protein.Titer of serum was determined by infrared scanning map,indirect ELISA and protein electrophoresis.Result：Titer of serum in the forth immunization was reached to 8 100,and competitive ratio was reached to 29.20%,which were respectively detected by the method of indirect ELISA and competitive inhibition ELISA.Conclusion：Although the serum has a little low titer,it has a high competitive ratio.It suggests that polyclonal antibody against elaidic acid is really exist in serum of immunization.This experiment could provide reliable experimental evidence and a theoretical basis for isolation and purification of polyclonal antibody against trans fatty acids.Furthermore,it will be a new approach to build an excellent experimental foundation for the detection of trans fatty acids in food by ELISA.