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Int. J. Mol. Sci. 2017, 18(7), 1507; doi:10.3390/ijms18071507

Rapid and Direct VHH and Target Identification by Staphylococcal Surface Display Libraries

BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schaenzlestrasse 18, 79104 Freiburg, Germany
Received: 22 June 2017 / Revised: 7 July 2017 / Accepted: 10 July 2017 / Published: 12 July 2017
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

Unbiased and simultaneous identification of a specific antibody and its target antigen has been difficult without prior knowledge of at least one interaction partner. Immunization with complex mixtures of antigens such as whole organisms and tissue extracts including tumoral ones evokes a highly diverse immune response. During such a response, antibodies are generated against a variety of epitopes in the mixture. Here, we propose a surface display design that is suited to simultaneously identify camelid single domain antibodies and their targets. Immune libraries of single-domain antigen recognition fragments from camelid heavy chain-only antibodies (VHH) were attached to the peptidoglycan of Gram-positive Staphylococcus aureus employing its endogenous housekeeping sortase enzyme. The sortase transpeptidation reaction covalently attached the VHH to the bacterial peptidoglycan. The reversible nature of the reaction allowed the recovery of the VHH from the bacterial surface and the use of the VHH in downstream applications. These staphylococcal surface display libraries were used to rapidly identify VHH as well as their targets by immunoprecipitation (IP). Our novel bacterial surface display platform was stable under harsh screening conditions, allowed fast target identification, and readily permitted the recovery of the displayed VHH for downstream analysis. View Full-Text
Keywords: VHH; nanobody; bacterial surface display; target identification; staphylococcal sortase A; immunoprecipitation VHH; nanobody; bacterial surface display; target identification; staphylococcal sortase A; immunoprecipitation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Cavallari, M. Rapid and Direct VHH and Target Identification by Staphylococcal Surface Display Libraries. Int. J. Mol. Sci. 2017, 18, 1507.

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