Journal
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
Volume 65, Issue 5, Pages 273-283Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1369/0022155417694869
Keywords
analyte; analytic response curve; HER-2; IHControl; immunohistochemistry; peptide
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Funding
- National Institutes of Health (NIH), National Cancer Institute [R44CA183203]
- National Center for Advancing Translational Sciences (also NIH) [UL1TR001064]
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An important limitation in the field of immunohistochemistry (IHC) is the inability to correlate stain intensity with specific analyte concentrations. Clinical immunohistochemical tests are not described in terms of analytic response curves, namely, the analyte concentrations in a tissue sample at which an immunohistochemical stain (1) is first visible, (2) increases in proportion to the analyte concentration, and (3) ultimately approaches a maximum color intensity. Using a new immunostaining tool (IHControls), we measured the analytic response curves of the major clinical immunohistochemical tests for human epidermal growth factor receptor type II (HER-2), estrogen receptor (ER), and progesterone receptor (PR). The IHControls comprise the analytes HER-2, ER, and PR at approximately log concentration intervals across the range of biological expression, from 100 to 1,000,000 molecules per test microbead. We stained IHControls of various concentrations using instruments, reagents, and protocols from three major IHC vendors. Stain intensity at each analyte concentration was measured, thereby generating an analytic response curve. We learned that for HER-2 and PR, there is significant variability in test results between clinical kits for samples with analyte concentrations of approximately 10(4) molecules/microbead. We propose that the characterization of immunostains is an important step toward standardization.
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