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Antigenic Characterization of Immunodominant Epitopes of Alpha-8 giardin in Giardia lamblia

WEI Chao-jun1, JIA Yan-juan1, XU Hui1, WU Ling1, LU Si-qi2, LI Yong-hong1*   

  1. 1 Gansu Center for Clinical Laboratory, Gansu Provincial Hospital, Gansu 730000, China;2 Department of Pathogen Biology, Capital Medical University, Beijing 100069, China
  • Online:2016-10-30 Published:2016-11-09

Abstract: Objective To identify the dominant epitopes of α-8 giardin in Giardia lamblia, and analyze their immunoreactivity. Methods Three dominant epitopes G1(7-17 aa), G2(30-40 aa) and G3(296-306 aa) were predicted for α-8 giardin by bioinformatic analysis using DNAstar, Biosun and CLUSTAL W softwares. Each epitope was conjugated to keyhole limpethemocyanin(KLH) and then used to immunize female Chinchilla rabbits via multi-point(8-10 points) subcutaneous injections on the back(0.5 mg conjugate/rabbit, 100 μl at each point). Then the rabbits were boosted(0.2 mg conjugate/rabbit) on days 14, 21 and 28 after the first immunization. Blood was collected prior to each immunization, and carotid artery blood(50 ml) was collected on day 7 after the last immunization. The antibody titer for anti-KLH-G1, anti-KLH-G2 and anti-KLH-G3 sera was determined with indirect ELISA. The reactivity of the three anti-sera with recominant giardin(rGiardin) was analyzed with Western blotting. Results The concentration of the purified conjugates KLH-G1, KLH-G2, and KLH-G3 was 0.66, 0.95 and 0.25 mg/ml, respectively. ELISA showed that the antibody titer for anti-KLH-G1, anti-KLH-G2 and anti-KLH-G3 sera was 1∶12800, 1∶51200, 1∶51200, respectively. SDS-PAGE revealed a specific band at Mr 36 000 for the three anti-sera. Western blotting showed that the three anti-sera all specifically recognized rGiardin. Conclusion Immunization with the KLH-G1, KLH-G2, and KLH-G3 conjugates induced production of specific IgG antibody in rabbits, which can recognize rGiardin.

Key words: Giardia lamblia, α-8 giardin, Bioinformatics analysis, Immunodominant epitopes