title: Development of a highly specific HER2 monoclonal antibody for immunohistochemistry using protein microarray chips.
abstract: HER2 is an orphan receptor tyrosine kinase of the EGFR families and is considered to be a key tumor driver gene [1]. Breast cancer and gastric cancer with HER2 amplification can be effectively treated by its neutralizing antibody, Herceptin. In clinic, Immunohistochemistry (IHC) was used as the primary screening method to diagnose HER2 amplification [2]. However, recent evidence suggested that the frequently used rabbit HER2 antibody 4B5 cross reacted with another family member HER4 [3]. IHC staining with 4B5 also indicated that there was strong non-specific cytoplasmic and nuclear signals in normal gastric mucosal cells and some gastric cancer samples. Using a protein lysate array which covers 85% of the human proteome, we have confirmed that the 4B5 bound to HER4 and a nuclear protein ZSCAN18 besides HER2. The non-specific binding accounts for the unexpected cytoplasmic and nuclear staining of 4B5 of normal gastric epithelium. Finally, we have developed a novel mouse HER2 monoclonal antibody UMAB36 with similar sensitivity to 4B5 but only reacted to HER2 across the 17,000 proteins on the protein chip. In 129 breast cancer and 158 gastric cancer samples, UMAB36 showed 100% sensitivity and specificity comparing to the HER2 FISH reference results with no unspecific staining in the gastric mucosa layer. Therefore, UMAB36 could provide as an alternative highly specific IHC reagent for testing HER2 amplification in gastric cancer populations.
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Lili
authors.initials: L
authors.lastname: Qi
authors.affiliation: Department of Pathology, Beijing Cancer Hospital, No. 52 Fu-Cheng Road, Haidian District, Beijing, 100142, PR China.
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Institute of Functional Nano and Soft Materials (FUNSOM), Collaborative Innovation Center of Suzhou Nano Science and Technology, Soochow University, Suzhou, 215123, PR China.
authors.forename: Mingmin
authors.initials: M
authors.lastname: Lu
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Kehu
authors.initials: K
authors.lastname: Yuan
authors.affiliation: Department of Pathology, Beijing Cancer Hospital, No. 52 Fu-Cheng Road, Haidian District, Beijing, 100142, PR China.
authors.forename: Zhongwu
authors.initials: Z
authors.lastname: Li
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Guiyin
authors.initials: G
authors.lastname: Wu
authors.affiliation: Department of Pathology, Beijing Cancer Hospital, No. 52 Fu-Cheng Road, Haidian District, Beijing, 100142, PR China.
authors.forename: Xiaozheng
authors.initials: X
authors.lastname: Huang
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Pathology, Beijing Cancer Hospital, No. 52 Fu-Cheng Road, Haidian District, Beijing, 100142, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhao
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Fu
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Boyang
authors.initials: B
authors.lastname: Chu
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA.
authors.forename: Guangli
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College of Soochow University, Suzhou, 215123, PR China. Electronic address: ffren@suda.edu.cn.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Ren
authors.affiliation: OriGene Technologies, 9620 Medical Center Dr., Suite 200, Rockville, MD, 20850, USA. Electronic address: dma@origene.com.
authors.forename: Donghui
authors.initials: D
authors.lastname: Ma
authors.affiliation: Institute of Functional Nano and Soft Materials (FUNSOM), Collaborative Innovation Center of Suzhou Nano Science and Technology, Soochow University, Suzhou, 215123, PR China. Electronic address: Chenjian2014@suda.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D003429:Cross Reactions; D006801:Humans; D007150:Immunohistochemistry; D040081:Protein Array Analysis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D013274:Stomach Neoplasms / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human
keywords: 4B5; HER2 amplification; Immunohistochemistry; UMAB36