Measuring tissue-based biomarkers by immunochromatography coupled with reverse-phase lysate microarray.

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
Martin J RomeoDavid M Berman

Abstract

There is a need for new technologies to study tissue-based biomarkers. The current gold standard, immunohistochemistry, is compromised by variability in tissue processing and observer bias. Reverse transcription-PCR (RT-PCR), immunocytochemistry, and reverse-phase lysate microarrays (RPM) are promising alternative technologies but have not yet been validated, or correlated, on the same patient-derived tissues. Furthermore, RPM is currently limited by time-consuming microdissection and low amounts of evaluable protein lysates. Metastatic melanoma was surgically excised from 30 patients and macroscopically dissected from surrounding stroma. Each specimen was processed by formalin-fixation (immunohistochemistry), cytospin (immunocytochemistry), or disaggreagation and enrichment (RT-PCR and RPM). The latter protocol uses immunochromatography to remove hematopoetic-derived cells, thus enriching for melanoma cells. Each sample was measured for the expression of gp100 or MART-1 normalized to actin. Immunochromatography coupled with RPM (I-RPM) is reproducible (r >/= 0.70) and, for gp100, correlates strongly with immunohistochemistry and immunocytochemistry (r = 0.78 and 0.76, respectively) and moderately with transcript levels, measur...Continue Reading

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Citations

Jul 23, 2008·The Analyst·Hye Jin LeeRobert M Corn
May 24, 2012·Journal of Biomedicine & Biotechnology·Robert Wellhausen, Harald Seitz
Feb 1, 2009·Proteomics. Clinical Applications·Robert J CaiazzoBrian C-S Liu
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Apr 30, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Timothy L FrankelRichard A Morgan

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