Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.

The identification of new markers, the expression of which defines new phenotipically and functionally distinct cell subsets, is a main objective in cell biology. We have addressed the issue of identifying new cell specific markers with a reverse proteomic approach whereby approximately 1700 human o...

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Autores principales:	Monica Moro, Mariacristina Crosti, Pasquale Creo, Pierangela Gallina, Serena Curti, Elisa Sugliano, Rossana Scavelli, Davide Cattaneo, Elena Canidio, Maurizio Marconi, Paolo Rebulla, Paolo Sarmientos, Giuseppe Viale, Massimiliano Pagani, Sergio Abrignani
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/17b80e11e6984370af5867ed8a6d98a1
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spelling	oai:doaj.org-article:17b80e11e6984370af5867ed8a6d98a12021-11-18T07:23:15ZIdentification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.1932-620310.1371/journal.pone.0034395https://doaj.org/article/17b80e11e6984370af5867ed8a6d98a12012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22496798/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The identification of new markers, the expression of which defines new phenotipically and functionally distinct cell subsets, is a main objective in cell biology. We have addressed the issue of identifying new cell specific markers with a reverse proteomic approach whereby approximately 1700 human open reading frames encoding proteins predicted to be transmembrane or secreted have been selected in silico for being poorly known, cloned and expressed in bacteria. These proteins have been purified and used to immunize mice with the aim of obtaining polyclonal antisera mostly specific for linear epitopes. Such a library, made of about 1600 different polyclonal antisera, has been obtained and screened by flow cytometry on cord blood derived CD34+CD45dim cells and on peripheral blood derived mature lymphocytes (PBLs). We identified three new proteins expressed by fractions of CD34+CD45dim cells and eight new proteins expressed by fractions of PBLs. Remarkably, we identified proteins the presence of which had not been demonstrated previously by transcriptomic analysis. From the functional point of view, looking at new proteins expressed on CD34+CD45dim cells, we identified one cell surface protein (MOSC-1) the expression of which on a minority of CD34+ progenitors marks those CD34+CD45dim cells that will go toward monocyte/granulocyte differentiation. In conclusion, we show a new way of looking at the membranome by assessing expression of generally neglected proteins with a library of polyclonal antisera, and in so doing we have identified new potential subsets of hematopoietic progenitors and of mature PBLs.Monica MoroMariacristina CrostiPasquale CreoPierangela GallinaSerena CurtiElisa SuglianoRossana ScavelliDavide CattaneoElena CanidioMaurizio MarconiPaolo RebullaPaolo SarmientosGiuseppe VialeMassimiliano PaganiSergio AbrignaniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 4, p e34395 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Monica Moro Mariacristina Crosti Pasquale Creo Pierangela Gallina Serena Curti Elisa Sugliano Rossana Scavelli Davide Cattaneo Elena Canidio Maurizio Marconi Paolo Rebulla Paolo Sarmientos Giuseppe Viale Massimiliano Pagani Sergio Abrignani Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
description	The identification of new markers, the expression of which defines new phenotipically and functionally distinct cell subsets, is a main objective in cell biology. We have addressed the issue of identifying new cell specific markers with a reverse proteomic approach whereby approximately 1700 human open reading frames encoding proteins predicted to be transmembrane or secreted have been selected in silico for being poorly known, cloned and expressed in bacteria. These proteins have been purified and used to immunize mice with the aim of obtaining polyclonal antisera mostly specific for linear epitopes. Such a library, made of about 1600 different polyclonal antisera, has been obtained and screened by flow cytometry on cord blood derived CD34+CD45dim cells and on peripheral blood derived mature lymphocytes (PBLs). We identified three new proteins expressed by fractions of CD34+CD45dim cells and eight new proteins expressed by fractions of PBLs. Remarkably, we identified proteins the presence of which had not been demonstrated previously by transcriptomic analysis. From the functional point of view, looking at new proteins expressed on CD34+CD45dim cells, we identified one cell surface protein (MOSC-1) the expression of which on a minority of CD34+ progenitors marks those CD34+CD45dim cells that will go toward monocyte/granulocyte differentiation. In conclusion, we show a new way of looking at the membranome by assessing expression of generally neglected proteins with a library of polyclonal antisera, and in so doing we have identified new potential subsets of hematopoietic progenitors and of mature PBLs.
format	article
author	Monica Moro Mariacristina Crosti Pasquale Creo Pierangela Gallina Serena Curti Elisa Sugliano Rossana Scavelli Davide Cattaneo Elena Canidio Maurizio Marconi Paolo Rebulla Paolo Sarmientos Giuseppe Viale Massimiliano Pagani Sergio Abrignani
author_facet	Monica Moro Mariacristina Crosti Pasquale Creo Pierangela Gallina Serena Curti Elisa Sugliano Rossana Scavelli Davide Cattaneo Elena Canidio Maurizio Marconi Paolo Rebulla Paolo Sarmientos Giuseppe Viale Massimiliano Pagani Sergio Abrignani
author_sort	Monica Moro
title	Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
title_short	Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
title_full	Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
title_fullStr	Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
title_full_unstemmed	Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
title_sort	identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/17b80e11e6984370af5867ed8a6d98a1
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Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins.

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