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Characterization of BRCAA1 and Its Novel Antigen Epitope Identification

¹ Bio-Nano-Engineering Center, Max Planck Institute for Metals Research, Stuttgart, Germany; ² Department of Dermatology of Xijing Hospital, ³ Department of Bioinformatics of Tangdu Hospital, ⁴ Institute of Genetics Diagnosis, Fourth Military Medical University, Xi'an, PR China; and ⁵ Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Lizuka, Japan

Daxiang Cui, Bio-Nano-Engineering Center, Max Planck Institute for Metals Research, Heisenbergstrasse 3, D-70569, Stuttgart, Germany. Phone: 49-711-689-3516; Fax: 49-711-689-3512. E-mail: dx.cui{at}mf.mpg.de

Looking for novel breast cancer antigen epitopes is helpful for its treatment, diagnosis, and prevention. brcaa1 gene is mapped at 1q42.1-q43, its whole genome is 93.857 kb, including 18 exons and 17 introns. BRCAA1 protein is composed of 1,214 amino acids with 10 glycosylate sites, and shares 37% amino acid identity and an identical antigen epitope with Rb binding protein 1. The novel antigen epitope, SSKKQKRSHK, was predicted to locate in the region 610 to 619 sites, was synthesized, and its antibody was fabricated. Competent inhibition analysis showed that SSKKQKRSHK is the shortest effective peptide. The antigen epitope was mapped in the cytoplasm of MCF-7 cells. Immunohistochemistry analysis showed that the antigen epitope exhibited positive expression in 65% (39 of 60) breast cancer specimens and negative expression in 60 non-cancerous tissues. Statistical analysis shows that its expression is closely associated with status of ER and PR, with sensitivity of 100% and specificity of 81%, and confidence interval of 85.9% to 96.9%. ELISA analysis showed that the mean absorbance of sera antibody titers from breast cancer patients and healthy donors were 0401 ± 0.163 SD and 0.137 ± 0.121 SD, respectively. Sixty-four percent breast cancer patient sera and 13% healthy donor sera had higher titer than mean titer of healthy donors, and there exists significant difference between breast cancer patients and healthy donors (P < 0.001). In this study, a novel breast cancer antigen epitope, SSKKQKRSHK, is identified. Its expression is associated with characteristics that are themselves associated with prognosis of breast cancer, and its sera antibody level may be helpful for breast cancer diagnosis.

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