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

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• 作者：Renata  ; Grifantini^a ;   ; ¹ ;   ; ^{renata.grifantini@externautics.com} ; Massimiliano  ; Pagani^b ;   ; ¹ ;   ; ² ; Andrea  ; Pierleoni^a ; Alberto  ; Grandi^a ; Matteo  ; Parri^a ; Susanna  ; Campagnoli^a ; Piero  ; Pileri^a ; Davide  ; Cattaneo^b ; Elena  ; Canidio^b ; Angela  ; Pontillo^b ; Elisa  ; De Camilli^c ; Alberto  ; Bresciani^b ; Federica  ; Marinoni^b ; Eros  ; Pedrazzoli^b ; Renzo  ; Nogarotto^a ; Sergio  ; Abrignani^d ; Giuseppe  ; Viale^f ; Paolo  ; Sarmientos^a ;   ; ^b ; Guido  ; Grandi^e
• 关键词：human proteins ; antibody collection ; immunohistochemistry ; tissue microarray
• 刊名：Journal of Proteomics
• 出版年：2011
• 出版时间：21 December 2011
• 年：2011
• 卷：75
• 期：2
• 页码：532-547
• 全文大小：2368 K

文摘

The YOMICS?antibody library () 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.