High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.

Monoclonal and recombinant antibodies are ubiquitous tools in diagnostics, therapeutics, and biotechnology. However, their biochemical properties lack optimal robustness, their bacterial production is not easy, and possibilities to create multifunctional fusion proteins based on them are limited. Mo...

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Autores principales:	Annika Järviluoma, Tomas Strandin, Sebastian Lülf, Jérôme Bouchet, Anna R Mäkelä, Matthias Geyer, Serge Benichou, Kalle Saksela
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f5d5bb140c814e85a52a5463f9b77ddf
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spelling	oai:doaj.org-article:f5d5bb140c814e85a52a5463f9b77ddf2021-11-18T07:13:20ZHigh-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.1932-620310.1371/journal.pone.0040331https://doaj.org/article/f5d5bb140c814e85a52a5463f9b77ddf2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22792285/?tool=EBIhttps://doaj.org/toc/1932-6203Monoclonal and recombinant antibodies are ubiquitous tools in diagnostics, therapeutics, and biotechnology. However, their biochemical properties lack optimal robustness, their bacterial production is not easy, and possibilities to create multifunctional fusion proteins based on them are limited. Moreover, the binding affinities of antibodies towards their antigens are suboptimal for many applications where they are commonly used. To address these issues we have made use of the concept of creating high binding affinity based on multivalent target recognition via exploiting some of the best features of immunoglobulins (Ig) and non-Ig-derived ligand-binding domains. We have constructed a small protein, named Neffin, comprised of a 118 aa llama Ig heavy chain variable domain fragment (VHH) fused to a ligand-tailored 57 aa SH3 domain. Neffin could be readily produced in large amounts (>18 mg/L) in the cytoplasm of E. coli, and bound with a subpicomolar affinity (K(d) 0.54 pM) to its target, the HIV-1 Nef protein. When expressed in human cells Neffin could potently inhibit Nef function. Similar VHH-SH3 fusion proteins could be targeted against many other proteins of interest and could have widespread use in diverse medical and biotechnology applications where biochemical robustness and strong binding affinity are required.Annika JärviluomaTomas StrandinSebastian LülfJérôme BouchetAnna R MäkeläMatthias GeyerSerge BenichouKalle SakselaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e40331 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Annika Järviluoma Tomas Strandin Sebastian Lülf Jérôme Bouchet Anna R Mäkelä Matthias Geyer Serge Benichou Kalle Saksela High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
description	Monoclonal and recombinant antibodies are ubiquitous tools in diagnostics, therapeutics, and biotechnology. However, their biochemical properties lack optimal robustness, their bacterial production is not easy, and possibilities to create multifunctional fusion proteins based on them are limited. Moreover, the binding affinities of antibodies towards their antigens are suboptimal for many applications where they are commonly used. To address these issues we have made use of the concept of creating high binding affinity based on multivalent target recognition via exploiting some of the best features of immunoglobulins (Ig) and non-Ig-derived ligand-binding domains. We have constructed a small protein, named Neffin, comprised of a 118 aa llama Ig heavy chain variable domain fragment (VHH) fused to a ligand-tailored 57 aa SH3 domain. Neffin could be readily produced in large amounts (>18 mg/L) in the cytoplasm of E. coli, and bound with a subpicomolar affinity (K(d) 0.54 pM) to its target, the HIV-1 Nef protein. When expressed in human cells Neffin could potently inhibit Nef function. Similar VHH-SH3 fusion proteins could be targeted against many other proteins of interest and could have widespread use in diverse medical and biotechnology applications where biochemical robustness and strong binding affinity are required.
format	article
author	Annika Järviluoma Tomas Strandin Sebastian Lülf Jérôme Bouchet Anna R Mäkelä Matthias Geyer Serge Benichou Kalle Saksela
author_facet	Annika Järviluoma Tomas Strandin Sebastian Lülf Jérôme Bouchet Anna R Mäkelä Matthias Geyer Serge Benichou Kalle Saksela
author_sort	Annika Järviluoma
title	High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
title_short	High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
title_full	High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
title_fullStr	High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
title_full_unstemmed	High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.
title_sort	high-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored sh3 domain.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/f5d5bb140c814e85a52a5463f9b77ddf
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High-affinity target binding engineered via fusion of a single-domain antibody fragment with a ligand-tailored SH3 domain.

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