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

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

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 1 2011
Externally publishedYes

Fingerprint

Flow cytometry
Mycobacterium
Sorting
Flow Cytometry
Clone Cells
Ovalbumin
Pathogens
Epitopes
Assays
Mucosal Immunity
Antigens
HIV-1
Population

ASJC Scopus subject areas

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

Cite this

Flow cytometry sorting of recombinant mycobacterial species yields bacterial clones with enhanced insert expression. / Yu, Jae Sung; Whitesides, John; Lee, Sunhee; Taylor, Natalie; Jacobs, William R.; Letvin, Norman L.; Haynes, Barton F.

In: Clinical and Vaccine Immunology, Vol. 18, No. 1, 01.01.2011, p. 43-49.

Research output: Contribution to journalArticle

Yu, Jae Sung ; Whitesides, John ; Lee, Sunhee ; Taylor, Natalie ; Jacobs, William R. ; Letvin, Norman L. ; Haynes, Barton F. / Flow cytometry sorting of recombinant mycobacterial species yields bacterial clones with enhanced insert expression. In: Clinical and Vaccine Immunology. 2011 ; Vol. 18, No. 1. pp. 43-49.
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