Enrichment of Native and Recombinant Extracellular Vesicles of MycobacteriaPraapti Jayaswal 1, Mohd Ilyas 1, Kuljit Singh 1,2, Saurabh Kumar 1,3, Lovely Sisodiya 1, Sapna Jain 1, Rahul Mahlawat 1, Nishant Sharma 1,4, Vishal Gupta 1, Krishnamohan Atmakuri 1
1Bacterial Pathogenesis Laboratory, Infectious Diseases and Immunology Group, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 2Clinical Microbiology Division, CSIR-Indian Institute of Integrative Medicine, 3ICAR-Research Complex for Eastern Region, 4Public Health Research Institute, Rutgers University
This protocol details the enrichment of native mycobacterial extracellular vesicles (mEVs) from axenic cultures of Mycobacterium smegmatis (Msm) and how mCherry (a red fluorescent reporter)-containing recombinant MsmEVs can be designed and enriched. Lastly, it verifies the novel approach with the enrichment of MsmEVs containing the EsxA protein of Mycobacterium tuberculosis.