运用蛋白质芯片技术鉴定干扰素的亚型

Identification of the type and the subtype of interferon with the application of protein biochips

目的:采用自制的6株干扰素单克隆抗体,在本实验室建立的金基底蛋白质芯片平台上对IFN-α1b、IFN-α2a、IFN-α2b、IFN-β及IFN-γ进行鉴定。方法:金基底蛋白质芯片用已知亚型的IFN半成品包被,封闭后与自制的6株干扰素单抗反应,再与Cy3-羊抗鼠IgC反应,反应后GenePix4100A芯片扫描仪扫描读数,将结果与ELISA法、Westernblot法比较。同样方法检测IFN成品。结果:金基底蛋白质芯片检测技术结果表明,该平台鉴定干扰素的亚型具有特异性,与ELISA法、Westernblot法能得出一致的结论。结论:运用本实验室建立的蛋白质芯片平台可以对干扰素的亚型进行鉴定,具有高通量,操作简单,样品用量少,重复性强等优点。

Objective:To establish a protein biochip flat to identify the subtype of interfon by adopting the 6 mAb which we have prepared.Methods: Recombinant human interferon was onto the NHS modified gold biochips.then the protein biochips were incubated with the 6 mAb.Cy3 conjugated sheep anti-mouse IgG was used to detect the antigen-antibody.Arrays are imaged using a fluorescence scanner (GenePix4100A) at 532 nm for Cy3.This method was compared with the traditional ones:ELISA,Western blot.Results:The results showed that this flat had speciality to identify the subtype of interfon. Conclusion: The protein biochip plat can provide a practical method to identify the subtype of interfon.It also has some advantages:high quantities;simple operation;low sample dosage and high repetition.