Application of immunoprecipitation-mass spectrometry for screening of antibody-recognizing molecules against Echinococcus granulosus
10.16303/j.cnki.1005-4545.2025.03.17
- VernacularTitle:应用免疫沉淀-质谱联用技术筛选细粒棘球绦虫抗体识别分子
- Author:
Xiaoxia WU
1
;
Jing DING
;
Xuemin JIN
;
Zhuangzhi ZHANG
;
Lixiao ZHANG
;
Mingyuan LIU
Author Information
1. 吉林大学动物医学学院/人畜共患传染病重症诊治全国重点实验室/人兽共患病研究教育部重点实验室,吉林长春 130062
- Publication Type:Journal Article
- Keywords:
hydatidosis;
Echinococcus granulosus;
immunoprecipitation-mass spectrometry(IP-MS);
antibody recognition molecule;
bioinformatics analysis
- From:
Chinese Journal of Veterinary Science
2025;45(3):519-526
- CountryChina
- Language:Chinese
-
Abstract:
We collected the positive serum of Echinococcus granulosus infection in sheep,an inter-mediate host with strong immune response,and used healthy serum as negative control,purified the serum and target protein to capture and enrich the corresponding antigen by immunoprecipita-tion,and obtained target protein-antibody-target protein complex.Mass spectrometry strategies were combined to screen and identify specific antigens associated with Echinococcus granulosus,and the proteins with the highest peptide coverage were analyzed bioinformatically using online prediction software.The results showed that 133 Echinococcus granulosus related proteins were i-dentified by IP-MS.Among them,one protein with peptide coverage≥70％was actin Ⅱ,and three proteins with peptide coverage between 30％to 40％were Ton B box domain containing protein,NADH dehydrogenase(ubiquinone)1 α-subcomplex 2(NADH dehydrogenase[ubiquinone])and lactic dehydrogenase.There were six proteins with 20％to 30％peptide coverage,namely,spli-cing factor 3B subunit 5,tumor protein D52,expressed conserved protein,NADH dehydrogenase(ubiquinone)1 alpha subcomplex 7,inosine-5'-monophosphate dehydrogenase,and aldo keto re-ductase family 1 member B4.Bioinformatics analysis revealed that actin protein has no signal pep-tide,it is probably a non-secretory protein and is subcellularly localized to the cytoskeleton,six op-timal potential antigenic epitopes are present,and the secondary and tertiary structures are consist-ently dominated by α-helices and irregular convolutions.The results indicate that immunoprecipita-tion-mass spectrometry is a high-throughput,simple,rapid and effective method for screening and identifying fine-grained Echinococcus granulosus antigens,which can provide a basis for screening specific molecules for serodiagnostic markers in intermediate host sheep and for the development of novel diagnostic techniques for hydatid diseases.
