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Construction and evaluation of an antibody phage display library targeting heparan sulfate

Heparan sulfate (HS) is a linear polysaccharide with high structural diversity. Different HS epitopes have been detected and localized using single chain variable fragment (scFv) antibodies from a ‘single pot’ phage display library containing a randomized complementarity determining region of the he...

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Autores principales: Damen, Lars A.A., van de Westerlo, Els M.A., Versteeg, Elly M.M., van Wessel, Thierry, Daamen, Willeke F., van Kuppevelt, Toin H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7329785/
https://www.ncbi.nlm.nih.gov/pubmed/32468289
http://dx.doi.org/10.1007/s10719-020-09925-z
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author Damen, Lars A.A.
van de Westerlo, Els M.A.
Versteeg, Elly M.M.
van Wessel, Thierry
Daamen, Willeke F.
van Kuppevelt, Toin H.
author_facet Damen, Lars A.A.
van de Westerlo, Els M.A.
Versteeg, Elly M.M.
van Wessel, Thierry
Daamen, Willeke F.
van Kuppevelt, Toin H.
author_sort Damen, Lars A.A.
collection PubMed
description Heparan sulfate (HS) is a linear polysaccharide with high structural diversity. Different HS epitopes have been detected and localized using single chain variable fragment (scFv) antibodies from a ‘single pot’ phage display library containing a randomized complementarity determining region of the heavy chain (CDR3). In this study, we created a new library containing anti-HS scFvs that all harbor a dp-38 heavy chain segment where the CDR3 region was engineered to contain the XBBXBX heparin binding consensus site (X = any amino acid, B = R, K or H). The library contained ~1.73 × 10(6) unique antibodies and was biopanned against HS from several sources. The selected antibodies were sequenced and chemically/immunohistologically characterized. A number of 67 anti-HS scFv antibodies were selected, of which 31 contained a XBBXBX CDR3 sequence. There was a clear preference for glycine at the first and proline at the fourth position of the CDR3. The sequence GZZP(R/K)X (Z = R, K or H, but may also contain N, S, or Q) was unusually overrepresented. Selected antibodies reacted with HS/heparin, but not with other glycosaminoglycans. Antibodies reacted differentially with respect to N-, 2-O, or 6-O-desulfated heparin preparations, and showed distinct topologies of HS epitopes in rat kidney sections. The library may be instrumental in the selection of a large pool of HS epitope-specific antibodies, and - since all antibodies differ only in their 6 amino acid CDR region - may be a tool for a rational design of antibodies recognizing specific HS sulfation patterns. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10719-020-09925-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-73297852020-07-07 Construction and evaluation of an antibody phage display library targeting heparan sulfate Damen, Lars A.A. van de Westerlo, Els M.A. Versteeg, Elly M.M. van Wessel, Thierry Daamen, Willeke F. van Kuppevelt, Toin H. Glycoconj J Original Article Heparan sulfate (HS) is a linear polysaccharide with high structural diversity. Different HS epitopes have been detected and localized using single chain variable fragment (scFv) antibodies from a ‘single pot’ phage display library containing a randomized complementarity determining region of the heavy chain (CDR3). In this study, we created a new library containing anti-HS scFvs that all harbor a dp-38 heavy chain segment where the CDR3 region was engineered to contain the XBBXBX heparin binding consensus site (X = any amino acid, B = R, K or H). The library contained ~1.73 × 10(6) unique antibodies and was biopanned against HS from several sources. The selected antibodies were sequenced and chemically/immunohistologically characterized. A number of 67 anti-HS scFv antibodies were selected, of which 31 contained a XBBXBX CDR3 sequence. There was a clear preference for glycine at the first and proline at the fourth position of the CDR3. The sequence GZZP(R/K)X (Z = R, K or H, but may also contain N, S, or Q) was unusually overrepresented. Selected antibodies reacted with HS/heparin, but not with other glycosaminoglycans. Antibodies reacted differentially with respect to N-, 2-O, or 6-O-desulfated heparin preparations, and showed distinct topologies of HS epitopes in rat kidney sections. The library may be instrumental in the selection of a large pool of HS epitope-specific antibodies, and - since all antibodies differ only in their 6 amino acid CDR region - may be a tool for a rational design of antibodies recognizing specific HS sulfation patterns. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10719-020-09925-z) contains supplementary material, which is available to authorized users. Springer US 2020-05-28 2020 /pmc/articles/PMC7329785/ /pubmed/32468289 http://dx.doi.org/10.1007/s10719-020-09925-z Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Original Article
Damen, Lars A.A.
van de Westerlo, Els M.A.
Versteeg, Elly M.M.
van Wessel, Thierry
Daamen, Willeke F.
van Kuppevelt, Toin H.
Construction and evaluation of an antibody phage display library targeting heparan sulfate
title Construction and evaluation of an antibody phage display library targeting heparan sulfate
title_full Construction and evaluation of an antibody phage display library targeting heparan sulfate
title_fullStr Construction and evaluation of an antibody phage display library targeting heparan sulfate
title_full_unstemmed Construction and evaluation of an antibody phage display library targeting heparan sulfate
title_short Construction and evaluation of an antibody phage display library targeting heparan sulfate
title_sort construction and evaluation of an antibody phage display library targeting heparan sulfate
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7329785/
https://www.ncbi.nlm.nih.gov/pubmed/32468289
http://dx.doi.org/10.1007/s10719-020-09925-z
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