基于表面等离子体共振技术用鸡蛋黄抗体 IgY测定人血清中转铁蛋白

采用表面等离子体共振(SPR)方法, 用鸡蛋黄抗体(IgY)取代传统免疫检测中哺乳动物抗体IgG作为识别分子偶联于CM5传感芯片上, 对人血清中的转铁蛋白进行了检测. 考察了IgY在传感芯片上的偶联条件及芯片的再生条件. 结果表明, 在pH=4.0, IgY浓度为100 μg/mL, 流速为5 μL/min的最佳偶联条件下, SPR响应信号和转铁蛋白浓度在50~500 ng/mL范围内具有良好的线性关系, 检出限为39.56 ng/mL, 对人血清样品检测的日间变异系数<8%, 日内变异系数<5%, 平均回收率为86.22% ~94.51%.

Determination of Transferrin in Human Serum Samples Using Chicken Egg Yolk Antibodies(IgY) by Surface Plasmon Resonance

Chicken egg yolk antibodies(IgY) instead of traditional mammalian antibodies were immobilized to a sensor chip CM5 for the detection of transferrin in human serum by surface plasmon resonance(SPR).The conditions for immobilizing chicken anti-human transferring antibodies and regeneration were studied.Chicken egg yolk antibodies were immobilized under the condition of a flow rate of 5 μL/min,pH=4.0 and the concentrationof 100 μg/mL.The good linear range was obtained over 50—500 ng/mL,the detection limit was 39.56 ng/mL.The determination of transferrin in human serum showed good reproduction and precision,intra-day CV was below 5%,and inter-day CV was below 8%,average recoveries ranged from 86.22% to 94.51%.