A novel polyclonal antibody library for expression profiling of poorly characterized, membrane and secreted human proteins

Renata Grifantini, Massimiliano Pagani, Andrea Pierleoni, Alberto Grandi, Matteo Parri, Susanna Campagnoli, Piero Pileri, Davide Cattaneo, Elena Canidio, Angela Pontillo, Elisa De Camilli, Alberto Bresciani, Federica Marinoni, Eros Pedrazzoli, Renzo Nogarotto, Sergio Abrignani, Giuseppe Viale, Paolo Sarmientos, Guido Grandi

Research output: Contribution to journalArticlepeer-review

Abstract

The YOMICS™ antibody library (http://www.yomics.com/) presented in this article is a new collection of 1559 murine polyclonal antibodies specific for 1287 distinct human proteins. This antibody library is designed to target marginally characterized membrane-associated and secreted proteins. It was generated against human proteins annotated as transmembrane or secreted in GenBank, EnsEMBL, Vega and Uniprot databases, described in no or very few dedicated PubMed-linked publications. The selected proteins/protein regions were expressed in E. coli, purified and used to raise antibodies in the mouse. The capability of YOMICS™ antibodies to specifically recognize their target proteins either as recombinant form or as expressed in cells and tissues was confirmed through several experimental approaches, including Western blot, confocal microscopy and immunohistochemistry (IHC). Moreover, to show the applicability of the library for biomarker investigation by IHC, five antibodies against proteins either known to be expressed in some cancers or homologous to tumor-associated proteins were tested on tissue microarrays carrying tumor and normal tissues from breast, colon, lung, ovary and prostate. A consistent differential expression in cancer was observed. Our results indicate that the YOMICS™ antibody library is a tool for systematic protein expression profile analysis that nicely complements the already available commercial antibody collections.

Original languageEnglish
Pages (from-to)532-547
Number of pages16
JournalJournal of Proteomics
Volume75
Issue number2
DOIs
Publication statusPublished - Dec 21 2011

Keywords

  • Antibody collection
  • Human proteins
  • Immunohistochemistry
  • Tissue microarray

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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