Whole-bacterium ribosome display selection for isolation of highly specific anti-Staphyloccocus aureus Affitins for detection- and capture-based biomedical applications.

Authors:
Stanimir Kambarev
Stanimir Kambarev
3Institut de Recherche en Santé de l'Université de Nantes INSERM U892 - CNRS 6299 CCINA Team 13: Nuclear Oncology Research Group: Engineering of Recognition
Petar Vukojicic
Petar Vukojicic
Stanford University
Irvine | United States
Nathalie Caroff
Nathalie Caroff
Laboratoire de Bactériologie-Virologie-Hygiène Hospitalière
France
Stephane Corvec
Stephane Corvec
Université de Nantes
Nantes | France
Barbara Mouratou
Barbara Mouratou
Biochemisches Institut der Universität Zürich
Switzerland

Biotechnol Bioeng 2019 Apr 14. Epub 2019 Apr 14.

CRCINA, INSERM, CNRS, Université d'Angers, Université de Nantes, Nantes, France.

Detection and capture methods using antibodies have been developed to ensure identification of pathogens in biological samples. Though antibodies have many attractive properties, they also have limitations and there are needs to expand the panel of available affinity proteins with different properties. Affitins, that we developed from the Sul7d proteins, are a solid class of affinity proteins, which can be used as substitutes to antibodies or to complement them. We report the generation and characterization of antibacterial Affitins with high specificity for Staphylococcus aureus. For the first time, ribosome display selections were carried out using whole-living-cell and naïve combinatorial libraries, which avoid production of protein targets and immunization of animals. We showed that Affitin C5 exclusively recognizes S. aureus among dozens of strains, including clinical ones. C5 binds staphylococcal Protein A (SpA) with a K of 108 ± 2 nM and has a high thermostability (T  = 77.0°C). Anti-S. aureus C5 binds SpA or bacteria in various detection and capture applications, including ELISA, western blot analysis, bead-fishing, and fluorescence imaging. Thus, novel anti-bacteria Affitins which are cost-effective, stable, and small can be rapidly and fully designed in vitro with high affinity and specificity for a surface-exposed marker. This class of reagents can be useful in diagnostic and biomedical applications.

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http://dx.doi.org/10.1002/bit.26989DOI Listing
April 2019

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References

(Supplied by CrossRef)
Expression of recombinant alkaline phosphatase conjugates in Escherichia coli
Boulain J. C. et al.
Methods in Molecular Biology 2004
Microfluidic sample preparation for medical diagnostics
Cui F. et al.
Annual Review of Biomedical Engineering 2015
Affitins for protein purification by affinity magnetic fishing
Fernandes C. S. M. et al.
Journal of Chromatography 2016
Development of aptamers against unpurified proteins
Goto S. et al.
Biotechnology and Bioengineering 2017
All individual domains of staphylococcal protein A show Fab binding
Jansson B. et al.
FEMS Immunology and Medical Microbiology 1998

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