吉林大学学报（医学版）2024，Vol.50Issue(2)：303-309,7.DOI:10.13481/j.1671-587X.20240202
抗单纯疱疹病毒1型的IgY制备及其生物学活性检测
Preparation of IgY against herpes simplex virus-1 and detection of its biological activity
摘要
Abstract
Objective:To prepare the anti-herpes simplex virus-1(HSV-1)egg yolk antibody(IgY)and discuss its biological activity,and to clarify its anti-HSV-1 capability.Methods:The inactivated HSV-1 vaccine was prepared,the high egg-producing hens were immunized by multi-site injection method in the chicken breast,and IgY was purified by polyethylene glycol-6000(PEG-6000)method;the titer,purity,and protein levels of IgY were detected by indirect ELISA method,sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)technology,and protein concentration quantification kits;the ability of IgY to neutralize the virus was detected by indirect ELISA method;the virus blocking rate was detected.The Vero cells were used as the target cells for viral infection.The cytopathic effect(CPE)at different antibody concentrations(0.015 60,0.031 25,0.062 50,0.125 00,0.250 00,0.500 00,and 1.000 00 g·L-1)was detected;the in vitro anti-HSV-1 capacity of IgY was detected according to the degrees of CPE.Results:The titer of the prepared IgY was gradually increased with the extension of immunization time.The titer of the IgY stabilized at 1/1 024 000;the clear bands for the IgY light and heavy chains were observed;the average protein level of IgY was 11.544 g·L-1;the blocking rate of IgY against HSV-1 could reach up to 77.90％;the degree of HSV-1 induced CPE was decreased with the increasing of the antibody concentrations,and when the concentration of IgY reached 0.500 00 g·L-1 or higher,less than 25％of the cells(even no cells)exhibited cytopathic changes.Conclusion:Anti-HSV-1 IgY is successfully prepared,and this antibody has significant neutralizing and blocking capabilities and in vitro inhibitory activity against HSV-1,which can be utilized for the development of drugs and diagnostic methods.关键词
单纯疱疹病毒1型/卵黄抗体/中和病毒/体外抗病毒能力Key words
Herpes simplex virus-1/Egg yolk antibody/Neutralizing virus/Antiviral ability in vitro分类
农业科技引用本文复制引用
苏海涛,翟玥,宋秀玲,徐坤..抗单纯疱疹病毒1型的IgY制备及其生物学活性检测[J].吉林大学学报（医学版）,2024,50(2):303-309,7.基金项目
吉林省科技厅科技创新基地(平台)建设项目(YDZJ202302CXJD054) （平台）
