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Selection of cell-type specific antibodies on tissue-sections using phage display

With the advent of modern technologies enabling single cell analysis, it has become clear that small sub-populations of cells or even single cells can drive the phenotypic appearance of tissue, both diseased and normal. Nucleic acid based technologies allowing single cell analysis has been faster to...

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Autores principales: Larsen, Simon Asbjørn, Meldgaard, Theresa, Lykkemark, Simon, Mandrup, Ole Aalund, Kristensen, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Ltd 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4549044/
https://www.ncbi.nlm.nih.gov/pubmed/25808085
http://dx.doi.org/10.1111/jcmm.12568
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author Larsen, Simon Asbjørn
Meldgaard, Theresa
Lykkemark, Simon
Mandrup, Ole Aalund
Kristensen, Peter
author_facet Larsen, Simon Asbjørn
Meldgaard, Theresa
Lykkemark, Simon
Mandrup, Ole Aalund
Kristensen, Peter
author_sort Larsen, Simon Asbjørn
collection PubMed
description With the advent of modern technologies enabling single cell analysis, it has become clear that small sub-populations of cells or even single cells can drive the phenotypic appearance of tissue, both diseased and normal. Nucleic acid based technologies allowing single cell analysis has been faster to mature, while technologies aimed at analysing the proteome at a single cell level is still lacking behind, especially technologies which allow single cell analysis in tissue. Introducing methods, that allows such analysis, will pave the way for discovering new biomarkers with more clinical relevance, as these may be unique for microenvironments only present in tissue and will avoid artifacts introduced by in vitro studies. Here, we introduce a technology enabling biomarker identification on small sub-populations of cells within a tissue section. Phage antibody libraries are applied to the tissue sections, followed by washing to remove non-bound phage particles. To eliminate phage antibodies binding to antigens ubiquitously expressed and retrieve phage antibodies binding specifically to antigens expressed by the sub-population of cells, the area of interest is protected by a ‘shadow stick’. The phage antibodies on the remaining areas on the slide are exposed to UV light, which introduces cross-links in the phage genome, thus rendering them non-replicable. In this work we applied the technology, guided by CD31 expressing endothelial cells, to isolate recombinant antibodies specifically binding biomarkers expressed either by the cell or in the microenvironment surrounding the endothelial cell.
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spelling pubmed-45490442015-08-28 Selection of cell-type specific antibodies on tissue-sections using phage display Larsen, Simon Asbjørn Meldgaard, Theresa Lykkemark, Simon Mandrup, Ole Aalund Kristensen, Peter J Cell Mol Med Original Articles With the advent of modern technologies enabling single cell analysis, it has become clear that small sub-populations of cells or even single cells can drive the phenotypic appearance of tissue, both diseased and normal. Nucleic acid based technologies allowing single cell analysis has been faster to mature, while technologies aimed at analysing the proteome at a single cell level is still lacking behind, especially technologies which allow single cell analysis in tissue. Introducing methods, that allows such analysis, will pave the way for discovering new biomarkers with more clinical relevance, as these may be unique for microenvironments only present in tissue and will avoid artifacts introduced by in vitro studies. Here, we introduce a technology enabling biomarker identification on small sub-populations of cells within a tissue section. Phage antibody libraries are applied to the tissue sections, followed by washing to remove non-bound phage particles. To eliminate phage antibodies binding to antigens ubiquitously expressed and retrieve phage antibodies binding specifically to antigens expressed by the sub-population of cells, the area of interest is protected by a ‘shadow stick’. The phage antibodies on the remaining areas on the slide are exposed to UV light, which introduces cross-links in the phage genome, thus rendering them non-replicable. In this work we applied the technology, guided by CD31 expressing endothelial cells, to isolate recombinant antibodies specifically binding biomarkers expressed either by the cell or in the microenvironment surrounding the endothelial cell. John Wiley & Sons, Ltd 2015-08 2015-03-26 /pmc/articles/PMC4549044/ /pubmed/25808085 http://dx.doi.org/10.1111/jcmm.12568 Text en © 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Larsen, Simon Asbjørn
Meldgaard, Theresa
Lykkemark, Simon
Mandrup, Ole Aalund
Kristensen, Peter
Selection of cell-type specific antibodies on tissue-sections using phage display
title Selection of cell-type specific antibodies on tissue-sections using phage display
title_full Selection of cell-type specific antibodies on tissue-sections using phage display
title_fullStr Selection of cell-type specific antibodies on tissue-sections using phage display
title_full_unstemmed Selection of cell-type specific antibodies on tissue-sections using phage display
title_short Selection of cell-type specific antibodies on tissue-sections using phage display
title_sort selection of cell-type specific antibodies on tissue-sections using phage display
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4549044/
https://www.ncbi.nlm.nih.gov/pubmed/25808085
http://dx.doi.org/10.1111/jcmm.12568
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