液相芯片技术抗体修饰制备条件的优化

Optimization of condition of suspension array antibody modified system

[目的]优化以琼脂糖水凝胶包裹的光子晶体为载体的液相芯片抗体修饰制备方法,提高抗体修饰密度及检测灵敏度。[方法]选取人Ig G作为待检靶蛋白,在琼脂糖水凝胶上首先固定羊抗人Ig G抗体,经过洗涤、封闭后,依次加入标准抗原、荧光标记的检测抗体,分别孵育、洗涤后,倒置荧光显微镜获取图像及荧光强度,根据荧光强度优化最佳琼脂糖水凝胶浓度、固定时间、氢氧化钠浓度、环氧氯丙烷浓度以及抗体浓度。[结果]在水凝胶浓度4%、Na OH浓度1.0mol/L、环氧氯丙烷浓度10%、反应时间3h、抗体浓度0.01mg/ml的条件下,荧光强度达到最强,即抗体修饰密度达到最大。[结论]该研究优化了以琼脂糖水凝胶包裹的光子晶体为载体的抗体修饰反应条件,提高了液相芯片检测的灵敏度。

[Objective]Optimization the condition of antibody modified to suspension array based on photonic crystal carrier which coating agarose gel,improved the density of antibodies on suspension array and detection sensitivity. [Methods]Taking human Ig G as the target protein to be inspected,in agarose gel we firstly fixed Goat antihuman Ig G antibody,after washing and closed,sequentially adding standard antigen and fluorescence labeled antibody,then incubated and washed,and used inverted fluorescence microscope to obtain images and detect fluorescence intensity. The best modified antibody agarose gel concentration,the optimum time,concentration of sodium hydroxide,epichlorohydrin concentration as well as the most suitable antibody concentration is determined according to the fluorescence intensity. [Results]The fluorescence intensity could reach the maximum under the optimum condition： 4% agarose gel,3 hours of reaction time,1. 0mol /L Na OH,10% epichlorohydrin and 0. 01 mg/ml antibody. The density of coating antibody could reach the maximum. [Conclusion]The study optimizating of antibody modification reaction conditions based on the photonic crystal agarose gel,improved the detection sensitivity of suspension array.