CAJ | 학술논문

基于竞争反应模式,建立了表面等离子体激元共振( Surface plasmon resonance, SPR)技术检测三聚氰胺的方法。首先在羧甲基葡聚糖修饰的芯片表面引入牛血清白蛋白-三聚氰胺偶联物( BSA-melamine),随后流动注射待测三聚氰胺与适量三聚氰胺单克隆抗体的混合溶液。基于溶液中三聚氰胺与芯片表面固定的BSA-melamine偶联物和溶液中固定浓度的三聚氰胺单克隆抗体之间的竞争反应,通过检测芯片表面结合的抗体所产生的SPR信号,对溶液中三聚氰胺进行定量分析。随着溶液中待测三聚氰胺浓度的增大,SPR信号随之降低。三聚氰胺检测的线性范围为0.25~13 nmol/L及13~250 nmol/L。通过流动注射NaOH溶液可实现芯片的再生,从而大大提高了样品分析通量。本方法解决了SPR技术不易直接检测低浓度小分子的缺陷,具有方便、快捷、成本低等优点。
기우경쟁반응모식,건립료표면등리자체격원공진( Surface plasmon resonance, SPR)기술검측삼취청알적방법。수선재최갑기포취당수식적심편표면인입우혈청백단백-삼취청알우련물( BSA-melamine),수후류동주사대측삼취청알여괄량삼취청알단극륭항체적혼합용액。기우용액중삼취청알여심편표면고정적BSA-melamine우련물화용액중고정농도적삼취청알단극륭항체지간적경쟁반응,통과검측심편표면결합적항체소산생적SPR신호,대용액중삼취청알진행정량분석。수착용액중대측삼취청알농도적증대,SPR신호수지강저。삼취청알검측적선성범위위0.25~13 nmol/L급13~250 nmol/L。통과류동주사NaOH용액가실현심편적재생,종이대대제고료양품분석통량。본방법해결료SPR기술불역직접검측저농도소분자적결함,구유방편、쾌첩、성본저등우점。
In this study, surface plasmon resonance ( SPR) assay of melamine was carried out based on a competitive reaction in which the assayed melamine with varying concentrations in solution competed with the pre-immobilized BSA-melamine conjugate for the anti-melamine monoclonal antibody ( mAb) in solution. The BSA-melamine conjugate was first immobilized on the carboxymethylated dextran-covered SPR chips, and this was followed by the injection of the mixed solution of melamine and anti-melamine mAb with a fixed concentration. By examining the SPR signals caused by the attached monoclonal antibodies, the concentration of melamine in solution could be determined. The linear range of the method was 0. 25-13 nmol/L and 13-250 nmol/L. The chips could be regenerated via injection of NaOH, thus, high-throughput assays could be attained. The developed method is convenient and cost-effective, holding great promise for direct quantification of small molecules in low quantities.