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bioRxiv · 10.1101/584615

Developing a routine lab test for absolute quantification of Her2 in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tissues using Quantitative Dot Blot (QDB) method

Yu, G.·Zhang, W.·Zhang, Y.·Lv, J.·Zhang, J.·Tang, F.
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Abstract

While the transition from current prevailing method of relative and semi-quantitative assessment of solid tumor biomarkers to their absolute quantitation promises improved objectivity, consistency and accuracy in daily clinical practice, this transition is hindered by lacking of suitable technique, especially for Formalin Fixed Paraffin Embedded (FFPE) tissue samples. In this retrospective study, Quantitative Dot Blot (QDB) method was adopted to measure Her2 levels absolutely and quantitatively at protein level using 2X15 m FFPE breast cancer tissue slices collected sequentially and non-selectively from local hospital. Her2 levels measured from 332 samples using two diagnostic antibodies respectively were strongly correlated (r=0.963, p=0.0000). When the result was evaluated with results from IHC or FISH analysis using Receiving Operating Characteristics (ROC) analysis, we were able to achieve 100% sensitivity (95% CI: 95.6% to 100%) and 99.3% specificity (95% CI: 96.14% to 99.98%) with results from IHC analysis using 0.267 nmole/g as cutoff, and 93.02% sensitivity (95%CI: 80.94% to 98.54%) and 93.68% specificity (95% CI: 86.76% to 97.65%) with results from FISH analysis using 0.261 nmole/g as cutoff. Thus, QDB method is demonstrated to provide objective and consistent assessment of Her2 levels in FFPE samples with comparable results from both IHC and FISH analyses. It is also the first method to achieve absolute quantitation of tissue biomarkers in FFPE samples to meet daily clinical need.

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Yu, G., Zhang, W., Zhang, Y., Lv, J., Zhang, J., Tang, F.. 2019-03-21. Developing a routine lab test for absolute quantification of Her2 in Formalin Fixed Paraffin Embedded (FFPE) breast cancer tissues using Quantitative Dot Blot (QDB) method. https://doi.org/10.1101/584615

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