This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wernérus, H. Articles by Ståhl, S. Search for Related Content PubMed PubMed Citation Articles by Wernérus, H. Articles by Ståhl, S. Agricola Articles by Wernérus, H. Articles by Ståhl, S.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, September 2003, p. 5328-5335, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5328-5335.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Fluorescence-Activated Cell Sorting of Specific Affibody-Displaying Staphylococci

Henrik Wernérus, Patrik Samuelson, and Stefan Ståhl^*

Department of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), SE-106 91 Stockholm, Sweden

Received 21 February 2003/ Accepted 26 June 2003

Efficient enrichment of staphylococcal cells displaying specific heterologous affinity ligands on their cell surfaces was demonstrated by using fluorescence-activated cell sorting. Using bacterial surface display of peptide or protein libraries for the purpose of combinatorial protein engineering has previously been investigated by using gram-negative bacteria. Here, the potential for using a gram-positive bacterium was evaluated by employing the well-established surface expression system for Staphylococcus carnosus. Staphylococcus aureus protein A domains with binding specificity to immunoglobulin G or engineered specificity for the G protein of human respiratory syncytial virus were expressed as surface display on S. carnosus cells. The surface accessibility and retained binding specificity of expressed proteins were demonstrated in whole-cell enzyme and flow cytometry assays. Also, affibody-expressing target cells could be sorted essentially quantitatively from a moderate excess of background cells in a single step by using a high-stringency sorting mode. Furthermore, in a simulated library selection experiment, a more-than-25,000-fold enrichment of target cells could be achieved through only two rounds of cell sorting and regrowth. The results obtained indicate that staphylococcal surface display of affibody libraries combined with fluoresence-activated cell sorting might indeed constitute an attractive alternative to existing technology platforms for affinity-based selections.

---

* Corresponding author. Mailing address: Department of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), SE-106 91 Stockholm, Sweden. Phone: 46 8 553 783 29. Fax: 46 8 553 784 81. E-mail: stefans{at}biotech.kth.se.

---

Applied and Environmental Microbiology, September 2003, p. 5328-5335, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5328-5335.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Kronqvist, N., Lofblom, J., Jonsson, A., Wernerus, H., Stahl, S. (2008). A novel affinity protein selection system based on staphylococcal cell surface display and flow cytometry. Protein Eng Des Sel 21: 247-255 [Abstract] [Full Text]  
• Lofblom, J., Sandberg, J., Wernerus, H., Stahl, S. (2007). Evaluation of Staphylococcal Cell Surface Display and Flow Cytometry for Postselectional Characterization of Affinity Proteins in Combinatorial Protein Engineering Applications. Appl. Environ. Microbiol. 73: 6714-6721 [Abstract] [Full Text]