Abstract 3216: Using protein microarray technology to develop mono-specific anti-ERCC1 mAbs for anatomic pathology applications

• Published in: Cancer research (Chicago, Ill.) Vol. 73; no. 8_Supplement; p. 3216
• Main Authors: Ma, Donghui, Shu, Youmin, Yuan, Kehu, He, Wei-Wu
• Format: Journal Article
• Language: English
• Published: 15-04-2013
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/1538-7445.AM2013-3216

Antibody with cross-reactivity can create unexpected side effects or false diagnostic reports if used for clinical purposes. ERCC1 is a predictive biomarker for cisplatin based chemotherapy. High ERCC1 expression is linked to drug resistance to cisplatin-based chemotherapy. 8F1 is one of the most commonly used monoclonal antibodies for evaluating ERCC1 expression levels in lung cancer patient tissue samples. By using high density protein microarray chip, we discovered that 8F1 not only reacts with its authentic target ERCC1, but also cross-reacts with an unrelated nuclear membrane protein, PCYT1A. The cross-reactivity to PCYT1A was further confirmed by other immunoassays, including Western blot, antigen absorption test, and immunohistochemistry (IHC) analyses. In this study, we also discovered that PCYT1A gene expression level is even significantly higher than ERCC1 gene expression level in a certain population of lung cancer patient tissue samples. In summary, 8F1 clone is not suitable for ERCC1 IHC assay due to its cross-reactivity with PCYT1A protein. To develop the best monoclonal antibody for ERCC1 IHC analysis, 18 monoclonal antibodies were generated and 6 of them were screened against our protein microarray chip. Two clones showed high mono-specificity on protein microarray chip test and both worked in IHC application.
Citation Format: Donghui Ma, Youmin Shu, Kehu Yuan, Wei-Wu He. Using protein microarray technology to develop mono-specific anti-ERCC1 mAbs for anatomic pathology applications. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3216. doi:10.1158/1538-7445.AM2013-3216