Analysis of protein expression in cell microarrays: a tool for antibody-based proteomics.
Tissue microarray (TMA) technology provides a possibility to explore protein expression patterns in a multitude of normal and disease tissues in a high-throughput setting. Although TMAs have been used for analysis of tissue samples, robust methods for studying in vitro cultured cell lines and cell aspirates in a TMA format have been lacking. We have adopted a technique to homogeneously distribute cells in an agarose gel matrix, creating an artificial tissue. This enables simultaneous profiling of protein expression in suspension- and adherent-grown cell samples assembled in a microarray. In addition, the present study provides an optimized strategy for the basic laboratory steps to efficiently produce TMAs. Presented modifications resulted in an improved quality of specimens and a higher section yield compared with standard TMA production protocols. Sections from the generated cell TMAs were tested for immunohistochemical staining properties using 20 well-characterized antibodies. Comparison of immunoreactivity in cultured dispersed cells and corresponding cells in tissue samples showed congruent results for all tested antibodies. We conclude that a modified TMA technique, including cell samples, provides a valuable tool for high-throughput analysis of protein expression, and that this technique can be used for global approaches to explore the human proteome.
Andersson
Ann-Catrin
AC
Department of Genetics and Pathology, Rudbeck Laboratory, University Hospital, Uppsala, Sweden.
Strömberg
Sara
S
Bäckvall
Helena
H
Kampf
Caroline
C
Uhlen
Mathias
M
Wester
Kenneth
K
Pontén
Fredrik
F
D000906:Antibodies / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D011506:Proteins / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D040901:Proteomics; D012680:Sensitivity and Specificity; D046888:Tissue Array Analysis / Q000379:methods*
D000906:Antibodies; D011506:Proteins