IngentaConnect Antibody-based tissue profiling as a tool for clinical proteomics

Authors: Kampf, Caroline¹; Andersson, Ann-Catrin²; Wester, Kenneth²; Björling, Erik²; Uhlen, Mathias³; Ponten, Fredrik²

Source: Clinical Proteomics, Volume 1, Numbers 3-4, September 2004 , pp. 285-299(15)

Publisher: Humana Press

Abstract:

Here, we show a strategy for high-throughput antibody-based tissue profiling with the aim to create an atlas of protein expression patterns in normal human tissues and cancer tissues representing the 20 most prevalent cancer types. A set of standardized tissue microarrays (TMAs) was produced to allow for rapid screening of a multitude of different cells and tissues using immunohistochemistry. Eight TMA blocks were produced containing 48 different normal human tissues in triplicate and cancer tissue from 216 individually different tumors in duplicate. Sections from these blocks were immunohistochemically stained using five commercial and five in-house generated antibodies. Digital images for annotation of expression profiles were generated using a semiautomated approach. Five hundred seventy-six images and annotation data corresponding to a total of 30 Gbytes of data were collected for each antibody. The data presented here suggest that antibody-based profiling of protein expression in tissues can be used as a valuable tool in clinical proteomics.

Keywords: Protein epitope signature tags; tissue microarray; protein atlas; antibody-based proteomics; monospecific antibodies

Document Type: Research article

DOI: 10.1385/CP:1:3-4:285

Affiliations: 1: Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85, Uppsala, Sweden, Email: caroline.kampf@genpat.uu.se 2: Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85, Uppsala, Sweden, 3: Department of Biotechnology, Royal Institute of Technology, Stockholm, Sweden,

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