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Updated: Aug 11, 2026
Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers
Published on: December 5, 2017
Quantitative immunohistochemistry and the use of cellular calibrators for HER2 receptor number determination
1GSK, Precision Medicine, Biomarker & Bioanalytical Platforms, Collegeville, PA, 19426, USA. Kevin.j.mckinski@gsk.com.
Insights
High error rates in immunohistochemistry (IHC) may be reduced by using cellular calibrators. This study details a method using cellular calibrators to quantify HER2 receptors in breast cancer, revealing analytical performance differences.
Area of Science:
- Biomedical Diagnostics
- Cancer Research
- Laboratory Medicine
Background:
- Immunohistochemistry (IHC) assays exhibit high error rates compared to other lab disciplines, often attributed to the absence of traceable analytical standards and quantitative controls.
- Current IHC workflows lack standardized calibrators, hindering accurate quantification and potentially impacting diagnostic reliability.
Purpose of the Study:
- To develop and evaluate a novel IHC method utilizing cellular calibrators for quantifying HER2 receptors in breast cancer tissues.
- To compare the analytical performance of IHC using cellular calibrators against established scoring methods.
Main Methods:
- Development of cellular calibrators with established HER2 receptor amounts using electrochemiluminescent immunoassay and flow cytometry.
- Application of the cellular calibrator-based IHC method to quantify HER2 in breast cancer tissue samples.
- Comparison of quantitated HER2 receptor numbers with standard 4B5 scoring results.
Main Results:
- The study successfully implemented cellular calibrators to determine the average number of HER2 receptors in breast cancer tumor cells.
- Significant discordance was observed between quantitated HER2 receptor numbers and the 4B5 scores, highlighting analytical performance variations.
- Cellular calibrators demonstrated potential for improving quantitative accuracy in IHC.
Conclusions:
- Cellular calibrators are ideal for IHC as they mimic endogenous antigens and can be processed identically to tissue samples.
- Novel methods for labeling cells with precise protein amounts are necessary for robust quantitative IHC.
- Development and validation of standards for quantitative IHC are crucial for demonstrating clinical utility and improving diagnostic accuracy.
Abstract:
It is suggested that high error rates in clinical IHC compared to other laboratory disciplines are due to a lack of traceable analytic standards with quantitative controls. Quantitative controls can be achieved by implementing calibrators into IHC workflows, but recently published approaches offer little discussion about what the ideal calibrator should look like. Here, we detail an IHC method that utilizes cellular calibrators to determine the average number of HER2 receptors in tumor cells of breast cancer tissue. Electrochemiluminescent immunoassay and flow cytometry were used to establish nominal HER2 amounts in the cellular calibrators. The quantitated number of HER2 receptors and associated 4B5 scores from commercially sourced breast cancer tumors were compared. The results were discordant, indicating differences in analytical performance. Cells are ideal calibrators for IHC because the antigens they express are structurally and functionally representative of the endogenous analyte being measured. They can also be stained and analyzed identically to tissues, and are easily cultured, expanded, and manipulated. However, novel methods for labeling cells with fixed amounts of proteins are needed. In addition, standards for validation of quantitative IHC methods should be developed and their clinical utility must be demonstrated.
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