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

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

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Article numbere34395
JournalPLoS One
Volume7
Issue number4
DOIs
Publication statusPublished - Apr 4 2012

Fingerprint

polyclonal antibodies
Antibodies
Lymphocytes
Blood
Proteins
proteins
antiserum
Immune Sera
cells
lymphocytes
blood
Cytology
Flow cytometry
surface proteins
granulocytes
transcriptomics
Fetal Blood
Granulocytes
cell biology
Computer Simulation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Moro, M., Crosti, M., Creo, P., Gallina, P., Curti, S., Sugliano, E., ... Abrignani, S. (2012). Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins. PLoS One, 7(4), [e34395]. https://doi.org/10.1371/journal.pone.0034395

Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins. / Moro, Monica; Crosti, Mariacristina; Creo, Pasquale; Gallina, Pierangela; Curti, Serena; Sugliano, Elisa; Scavelli, Rossana; Cattaneo, Davide; Canidio, Elena; Marconi, Maurizio; Rebulla, Paolo; Sarmientos, Paolo; Viale, Giuseppe; Pagani, Massimiliano; Abrignani, Sergio.

In: PLoS One, Vol. 7, No. 4, e34395, 04.04.2012.

Research output: Contribution to journalArticle

Moro, M, Crosti, M, Creo, P, Gallina, P, Curti, S, Sugliano, E, Scavelli, R, Cattaneo, D, Canidio, E, Marconi, M, Rebulla, P, Sarmientos, P, Viale, G, Pagani, M & Abrignani, S 2012, 'Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins', PLoS One, vol. 7, no. 4, e34395. https://doi.org/10.1371/journal.pone.0034395
Moro, Monica ; Crosti, Mariacristina ; Creo, Pasquale ; Gallina, Pierangela ; Curti, Serena ; Sugliano, Elisa ; Scavelli, Rossana ; Cattaneo, Davide ; Canidio, Elena ; Marconi, Maurizio ; Rebulla, Paolo ; Sarmientos, Paolo ; Viale, Giuseppe ; Pagani, Massimiliano ; Abrignani, Sergio. / Identification of new hematopoietic cell subsets with a polyclonal antibody library specific for neglected proteins. In: PLoS One. 2012 ; Vol. 7, No. 4.
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