共面多氯联苯抗体的制备及其用表面等离子共振方法检测的应用研究

Preparation of antibodies against coplanar polychlorinated biphenyls (Co-PCBs) and their application in surface plasmon resonance(SPR)

根据文献方法获得抗共面多氯联苯(Co-PCBs)的重组抗体的轻链、重链基因序列,并用柔性连接肽(Gly4Ser)3进行连接.然后将获得的重组抗体基因序列通过克隆、表达、纯化、复性等手段得到纯度大于95%的Co-PCBs抗体.最后将Co-PCBs抗体组装到金膜表面,制备成检测用的蛋白芯片,并应用表面等离子共振技术(SPR)对共面多氯联苯中的PCB126进行了检测.结果显示,得到的Co-PCBs抗体特异性强,而且SPR检测方法测定PCB126的检测限为5×10-4pg·L-1,达到环境中Co-PCBs的超痕量检测水平要求.整个检测过程方便快捷、样品用量少、灵敏度高,可为环境中多氯联苯的实时检测研究提供指导.

To construct the gene sequence of a recombinant antibody,the light and heavy gene sequences of recombinant antibodies against coplanar polychlorinated biphenyls （Co-PCBs） were indexed and connected with a flexible linker peptide （Gly4Ser）3,then cloned and expressed in Escherichia coli M15. After purifying and renaturation,the purity of the recombinant antibody against Co-PCBs was more than 95%. Finally the antibody was assembled on a gold surface to prepare a protein chip for detecting PCB126 basing on SPR technology. The detection limit was about 5×10-4 pg·L-1,which meets the need of ultra trace detection of Co-PCBs in the environment. This detection method was fast and convenient,and less sample is needed because of its high-sensitivity. The work provides a good foundation for the real-time detection of PCBs in the environment.