摘要
目的比较海藻糖(trehalose)以及其他封闭液在多种抗原抗体反应体系中以及膜基质反应体系中应用的可行性,并观察海藻糖对预包被板抗原、抗体稳定性的保护作用。方法采用间接、双抗体夹心ELISA、Dot-ELISA和Western Blot观察海藻糖作为封闭剂的作用,特别是对糖结合抗原抗体反应体系敏感性与检测本底的影响。结果在一定的时间内,不同浓度的海藻糖封闭液均可以在不影响各反应体系敏感性的前提下,抑制Dot-ELISA和Western Blot体系中的非特异性吸附,封闭效果明显优于其他二糖封闭液与部分传统封闭液;并可有效封闭糖结合抗原的抗体反应;更重要的是,海藻糖封闭液可在20天内有效保护37℃以上高温环境中预包被板蛋白的活性。结论海藻糖能够作为封闭液,应用于糖结合抗原抗体反应体系、部分以PVDF膜为基质的抗原抗体反应体系,并可以在一定时间内保护高温条件下预包被板抗原、抗体的稳定性。
Objective To canpare the blocking effect of trehalose with other blocking reagents in in different antigen-antibody reactions and PVDF membrane based reaction. And observe the capability of trehalose in keeping stability of pre-coating antigen and antibody. Methods Trehalose blocking reagent in different concerntration and blocking time in the direct, indirect, sandwich-ELISA, saccharide-conjugated antigen-antibody reaction and PVDF based reaction systerm were tested to observe the influence on sensitivity and no-specific absorption comparing with other disaccharides blocking reagent and some conventional blocking reagents. Results Trehalose blocking buffers of different concemtration are all able to decrease no-specific absorption while without decrease of sensitivity in each systerm,especiaUy in the systerm of carbohysrate-binding antigen-antibody reaction, which are much better than other disaccharide and some conventional blocking reagents.In addition, trehalose blocking reagent exhibits the protecting the activity of pre-coated microplate from degradation induced by temperature above 37℃ for 20 days. Conelusion Trehalose can be used in ELISA for carbohydrate-binding antigen-antibody reaction and Dot-ELISA and Western-Blot based on PVDEF, and also in keeping stability of pre-coating protein.
出处
《中国实验诊断学》
2007年第6期717-721,共5页
Chinese Journal of Laboratory Diagnosis
基金
广州市科技计划项目:06A1212008
