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Analytical Validation of a Highly Quantitative, Sensitive, Accurate, and Reproducible Assay (HERmark) for the Measurement of HER2 Total Protein and HER2 Homodimers in FFPE Breast Cancer Tumor Specimens.

Larson JS, Goodman LJ, Tan Y, Defazio-Eli L, Paquet AC, Cook JW, Rivera A, Frankson K, Bose J, Chen L, Cheung J, Shi Y, Irwin S, Kiss LD, Huang W, Utter S, Sherwood T, Bates M, Weidler J, Parry G, Winslow J, Petropoulos CJ, Whitcomb JM - Patholog Res Int (2010)

Bottom Line: We report here the results of the analytical validation of assays that measure HER2 total protein (H2T) and HER2 homodimer (H2D) expression in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tumors as well as cell line controls.The accuracy of H2T measurements spans a broad dynamic range (2-3 logs) as evaluated by comparison with cross-validating technologies.The measurement of H2T expression demonstrates a sensitivity that is approximately 7-10 times greater than conventional immunohistochemistry (IHC) (HercepTest).

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Laboratory Operations, Monogram Biosciences, Inc., South San Francisco, CA 94080, USA.

ABSTRACT
We report here the results of the analytical validation of assays that measure HER2 total protein (H2T) and HER2 homodimer (H2D) expression in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tumors as well as cell line controls. The assays are based on the VeraTag technology platform and are commercially available through a central CAP-accredited clinical reference laboratory. The accuracy of H2T measurements spans a broad dynamic range (2-3 logs) as evaluated by comparison with cross-validating technologies. The measurement of H2T expression demonstrates a sensitivity that is approximately 7-10 times greater than conventional immunohistochemistry (IHC) (HercepTest). The HERmark assay is a quantitative assay that sensitively and reproducibly measures continuous H2T and H2D protein expression levels and therefore may have the potential to stratify patients more accurately with respect to response to HER2-targeted therapies than current methods which rely on semiquantitative protein measurements (IHC) or on indirect assessments of gene amplification (FISH).

No MeSH data available.


Related in: MedlinePlus

Section size linearity of the H2T and H2D assays.  (a) Cell line controls with varying levels of H2T (a) and H2D (b) were sub-sectioned and then run in the HERmark assay to determine linearity of the measurements with respect to section size.  Normalized values (y-axis) show the final signals that have been corrected for size.  95.8% of the pairwise comparisons were within 1.7-fold in the H2T assay and 95% of the pairwise comparisons were with 2.2-fold for the H2D assay.
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fig7: Section size linearity of the H2T and H2D assays. (a) Cell line controls with varying levels of H2T (a) and H2D (b) were sub-sectioned and then run in the HERmark assay to determine linearity of the measurements with respect to section size. Normalized values (y-axis) show the final signals that have been corrected for size. 95.8% of the pairwise comparisons were within 1.7-fold in the H2T assay and 95% of the pairwise comparisons were with 2.2-fold for the H2D assay.

Mentions: For the H2T assays, 100% of the pairwise comparisons were within 2-fold and 95.8% of the pairwise comparisons were within 1.7-fold (Figure 7(a)). For the H2D assay, 95% of the pairwise comparisons were within 2.2-fold (Figure 7(b)).


Analytical Validation of a Highly Quantitative, Sensitive, Accurate, and Reproducible Assay (HERmark) for the Measurement of HER2 Total Protein and HER2 Homodimers in FFPE Breast Cancer Tumor Specimens.

Larson JS, Goodman LJ, Tan Y, Defazio-Eli L, Paquet AC, Cook JW, Rivera A, Frankson K, Bose J, Chen L, Cheung J, Shi Y, Irwin S, Kiss LD, Huang W, Utter S, Sherwood T, Bates M, Weidler J, Parry G, Winslow J, Petropoulos CJ, Whitcomb JM - Patholog Res Int (2010)

Section size linearity of the H2T and H2D assays.  (a) Cell line controls with varying levels of H2T (a) and H2D (b) were sub-sectioned and then run in the HERmark assay to determine linearity of the measurements with respect to section size.  Normalized values (y-axis) show the final signals that have been corrected for size.  95.8% of the pairwise comparisons were within 1.7-fold in the H2T assay and 95% of the pairwise comparisons were with 2.2-fold for the H2D assay.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2990097&req=5

fig7: Section size linearity of the H2T and H2D assays. (a) Cell line controls with varying levels of H2T (a) and H2D (b) were sub-sectioned and then run in the HERmark assay to determine linearity of the measurements with respect to section size. Normalized values (y-axis) show the final signals that have been corrected for size. 95.8% of the pairwise comparisons were within 1.7-fold in the H2T assay and 95% of the pairwise comparisons were with 2.2-fold for the H2D assay.
Mentions: For the H2T assays, 100% of the pairwise comparisons were within 2-fold and 95.8% of the pairwise comparisons were within 1.7-fold (Figure 7(a)). For the H2D assay, 95% of the pairwise comparisons were within 2.2-fold (Figure 7(b)).

Bottom Line: We report here the results of the analytical validation of assays that measure HER2 total protein (H2T) and HER2 homodimer (H2D) expression in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tumors as well as cell line controls.The accuracy of H2T measurements spans a broad dynamic range (2-3 logs) as evaluated by comparison with cross-validating technologies.The measurement of H2T expression demonstrates a sensitivity that is approximately 7-10 times greater than conventional immunohistochemistry (IHC) (HercepTest).

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Laboratory Operations, Monogram Biosciences, Inc., South San Francisco, CA 94080, USA.

ABSTRACT
We report here the results of the analytical validation of assays that measure HER2 total protein (H2T) and HER2 homodimer (H2D) expression in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tumors as well as cell line controls. The assays are based on the VeraTag technology platform and are commercially available through a central CAP-accredited clinical reference laboratory. The accuracy of H2T measurements spans a broad dynamic range (2-3 logs) as evaluated by comparison with cross-validating technologies. The measurement of H2T expression demonstrates a sensitivity that is approximately 7-10 times greater than conventional immunohistochemistry (IHC) (HercepTest). The HERmark assay is a quantitative assay that sensitively and reproducibly measures continuous H2T and H2D protein expression levels and therefore may have the potential to stratify patients more accurately with respect to response to HER2-targeted therapies than current methods which rely on semiquantitative protein measurements (IHC) or on indirect assessments of gene amplification (FISH).

No MeSH data available.


Related in: MedlinePlus