Expression and Immunogenicity Analysis of Recombinant Leptospira Interrogans Surface Protein LigA in Mouse Model.

• Tác giả: Aida Chalesh, Pejvak Khaki, Soheila Moradi Bidhendi, Morteza Taghizadeh Tarnabi, Majid Tebianian
• Ngôn ngữ: eng
• Ký hiệu phân loại: 664.72273 Grains, other seeds, their derived products
• Thông tin xuất bản: England : Veterinary medicine and science , 2025
• Bộ sưu tập: NCBI
• ID: 747554
• Chủ đề: 1. Expression; 2. And; 3. Immunogenicity

Tóm tắt

BACKGROUND: Pathogenic strains of spirochetes of Leptospira spp. cause a globally distributed zoonotic disease called leptospirosis. The disease has several clinical manifestations, ranging from asymptomatic and subclinical infection to fatal and severe forms. HYPOTHESIS/OBJECTIVES: The aim of this study was to produce a recombinant Leptospiral immunoglobulin-like surface protein-A (r-LigA) antigen of Leptospira interrogans in a prokaryotic expression system and to assess its efficacy in a mouse model. MATERIALS AND METHODS: The optimal epitopes of the LigA protein were identified via bioinformatics studies. The pET32a RESULTS: Bioinformatics analysis identified the key antigenic region of LigA spanning amino acids 852 to 1210. Colony PCR and digestion confirmed the successful transformation. Induction using 0.5 mM IPTG at 30°C for 5 h was found to be optimal. Overexpression of r-LigA under optimised conditions accumulated proteins as inclusion bodies. Purification of r-LigA under native conditions using optimised Ni-NTA yielded 1050 µg/mL protein and high immunogenicity by effectively stimulating the immune system in female Balb/C mice. CONCLUSIONS: These findings support r-LigA as a strong candidate for future leptospirosis diagnostic tools and subunit vaccine development.