摘要
为了筛选抗原封闭液中聚乙二醇(PEG4000)的适宜浓度,首先用狂犬病病毒(Rabies virus,RV)细胞培养物包被抗原板,以含1%BSA和5%蔗糖的PBST溶液中加入不同浓度的PEG作为封闭液,分别于4℃、37℃保存7,14,30,90,180 d,从中筛选含适宜浓度PEG的封闭液,然后分别用乙肝表面抗原(HBsAg)、猪瘟病毒E2抗原(CSFV E2)和猪繁殖与呼吸综合征nsp2基因(PRRSV nsp2)原核表达产物作为包被抗原,对所筛选出的封闭液进行稳定性评价。结果表明:当封闭液为含12%PEG+1%BSA+5%蔗糖的PBST时,上述4种抗原包被板均可在37℃稳定保存180 d。由于该封闭液对不同来源抗原所具有的良好稳定性,初步推断该封闭液可能具有较广泛的应用前景。
To screen the suitable concentration of polyethylene glycol 4000 （PEG4000） in a blocking solution of coated antigens. Firstly, the cell cultures of Rabies virus （RV） were used to coat ELISA plates with antigen, and the phosphate -buffered saline with Triton X - 100 （PBST） solutions containing 1% BSA and 5% sucrose with different concentrations of PEG 4000 used as blocking solutions, were stored at 4 ℃ or 37 ℃ for 7 d, 14 d,30 d,90 d, and 180 d, respectively; and then the suitable concentration of PEG in the blocking solution was screened out. Second- ly ,the screened blocking solution was evaluated for its stability using hepatic B surface antigen （HBsAg） ,classical swine fever virus E2 （ CSFV E2 ） antigen and prokaryotic expression products of porcine reproductive and respiratory syndrome virus nsp2 as the coated antigens,respective- ly. The results showed that when PBST containing 12% PEG and 1% BSA with 5% sucrose was exploited as a blocking solution, the four kinds of the antigen - coated plates could be stably stored at 37℃ for 180 d. It is prehminarily concluded that this blocking solution may have wide application prospects due to its excellent stability to the antigens from different sources.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2014年第10期6-9,共4页
Heilongjiang Animal Science And veterinary Medicine
基金
国家自然科学基金项目(31072147
31272579)
