摘要
目的探讨pH值对胶体金标记单克隆抗体（mAb）性能的影响。方法用K2CO3调整胶体金溶液的pH值,得到pH值在5.0-9.0范围内的胶体金溶液,并用抗肠道病毒71型（EV71）VP1 mAb进行标记,牛血清白蛋白（BSA）封闭。用紫外-可见光谱扫描监测该标记过程中胶体金的性能改变。采用Mey氏稳定化实验检测免疫金的稳定性并检测制得的成品胶体金试纸条的灵敏度。结果 EV71-VP1 mAb在胶体金溶液pH值为7.0-8.5范围内制备的免疫金较稳定,且制得试纸条灵敏度高。结论 pH值是影响胶体金稳定性、决定胶体金与mAb间作用力的重要因素,并证实了紫外-可见光谱扫描评价pH值对胶体金标记的效果,建立了用紫外-可见光谱选择最适标记pH的方法标准。
Objective To investigate the influence of pH on the properties of colloidal gold labeling monoclonal antibody（ mAb）. Methods The pH value of colloidal gold was adjusted by K2CO3 solution. Then colloidal gold with a range of pH 5. 0- 9. 0 labeled enterovirus 71（ EV71）-VP1 mAb respectively,and bovine serum albumin（ BSA） was added to block the unreacted sites on the gold colloids for further experiments. The changes in the properties of colloidal gold in the progress of colloidal gold labeling mAb were monitored by UV /Vis spectroscopy. Finally,Mey＇s test was adopted to identify the stability of immunogold,and the sensitivity of the strip was evaluated by detecting gradual dilution of serum. Results EV71-VP1 mAb could be conjugated with colloidal gold at pH7. 0- 8. 5,and at this optimal pH,the test strip presented a good sensitivity.Conclusion pH is an important factor to ensure the stability of immunogold and to determine the conjugation effect between colloidal gold and mAb. The study confirmed that UV /Vis spectroscopy can evaluate the influence of pH on the properties of colloidal gold labeling mAb and set up an optimal pH for colloidal gold labeling using UV /Vis spectroscopy.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第11期1170-1173,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
河南省科技厅重点攻关项目(102102310093)
关键词
胶体金
PH值
单克隆抗体
免疫金
colloidal gold
pH
monoclonal antibody
immunogold
