检验医学与临床2018，Vol.15Issue(2)：202-205,208,5.DOI:10.3969/j.issn.1672-9455.2018.02.018
抗铜绿假单胞菌IgY的制备优化及抑菌效果
The optimized preparation of IgY against p .aeruginosa and the detection of its antibiotic effect
姚羽菲 1龙敏 2文荟淋 2姚刚3
作者信息
- 1. 四川大学华西临床医学院 ,成都610041
- 2. 四川大学生命科学学院 ,成都610065
- 3. 成都安蒂康生物科技有限公司610041
- 折叠
摘要
Abstract
Objective To prepare the pure and effective immunoglobulin yolk (IgY ) against pseudomonas aeruginosa ,and optimize the preparation process .Methods Using these methods ,such as ultrasonication ,res-onance cracking ,grinding ,we lysed p .aeruginosa and gained the optimized lysis antigen .The laying hens were immunized with lysis antigen or inactivated bacteria .Indirect Enzyme-linked immunosorbent was used to eval-uate the titer and stability of IgY .Then IgY was isolated and purified from egg yolks with high-titer using wa-ter dilution-salting out method or ultrafiltration .The purity and concentration of IgY were analysed by SDS-PAGE and BCA ,respectively .The antibacterial effect was evaluated by plate method ,and the cell morphology was detected by scanning electron microscopy .Results Ultrasonication was the most effective way to crack the antigen of p .aeruginosa .Immunization scheme that 0 .5 mL inactivate bacteria (1 × 109 CFU/mL) were in-jected hens and secondly injected after 4 weeks was proved the best for the titer of 1:25600 .The purity of IgY was 94％ after ammonium sulfate precipitation and ultrafiltration purification and the concentration 27 .02 mg/mL ,2 .7 mg IgY could restrain the grow th of bacteria obviously and the colony number decreased 9/10 compared to the control group .Meanwhile ,the green pigment produced by p .aeruginosa was inhibited obviously .Scanning electron microscope showed p .aeruginosa were bigger and shrinking after treated with IgY .Conclusion T he preparation and purification IgY against P .aeruginosa has been successfully optimized . And the IgY antibody has specific and marked antibacterial activity .关键词
铜绿假单胞菌/鸡卵黄抗体/抑菌效应Key words
pseudomonas aeruginosa/immunoglobubin of yolk/antibiotic effect分类
生物科学引用本文复制引用
姚羽菲,龙敏,文荟淋,姚刚..抗铜绿假单胞菌IgY的制备优化及抑菌效果[J].检验医学与临床,2018,15(2):202-205,208,5.
