Format

Send to

Choose Destination
Biotechnol Bioeng. 2019 Aug;116(8):1844-1855. doi: 10.1002/bit.26989. Epub 2019 May 15.

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

Author information

1
CRCINA, INSERM, CNRS, Université d'Angers, Université de Nantes, Nantes, France.
2
EA3826 Thérapeutiques cliniques et expérimentales des infections, UFR de Médecine, Université de Nantes, Nantes, France.
3
CRCINA, INSERM, Université d'Angers, Université de Nantes, Nantes, France.
4
Service de Bactériologie - Hygiène hospitalière, CHU de Nantes, Nantes, France.

Abstract

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 D of 108 ± 2 nM and has a high thermostability (T m  = 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.

KEYWORDS:

Affitin; Protein A; Staphylococcus aureus; Sul7d; immuno-detection; ribosome display

PMID:
30982947
DOI:
10.1002/bit.26989

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center