Flow cytometry sorting of recombinant mycobacterial species yields bacterial clones with enhanced insert expression

Jae Sung Yu, John Whitesides, Sun Hee Lee, Natalie Taylor, William R. Jacobs, Norman L. Letvin, Barton F. Haynes

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Recombinant mycobacteria hold promise as vectors for delivery of HIV-1 and other pathogen antigen inserts for inducing systemic and mucosal immune responses. In general, the immunogenicity of the recombinant mycobacterial insert is proportional to the level of insert expression. In this study, a novel flow cytometry-based assay has been developed to sort live recombinant mycobacterial mutants with high expression of foreign inserts and to enrich those sorted bacterial populations. Sorted recombinant mycobacterial clones expressed higher levels of the ovalbumin SIINFEKL epitope, and select sorted clones showed better immunogenicity than unsorted recombinant mycobacteria. Thus, flow cytometry-based sorting can isolate recombinant mycobacteria enriched for higher insert expression.

Original languageEnglish (US)
Pages (from-to)43-49
Number of pages7
JournalClinical and Vaccine Immunology
Volume18
Issue number1
DOIs
StatePublished - Jan 2011

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

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