Chavany Christine, Dea Jeffrey, Hernández González Rafael, Valle Rosaura P C, Jendoubi Moncef
Milagen, Inc., 1255 Park Avenue, Suite B, Emeryville, CA, 94608, USA.
Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga 15, Belisario Domínguez Secc 16, Tlalpan, 14080, Ciudad de México, CDMX, Mexico.
Biochem Biophys Rep. 2025 Jun 20;43:102097. doi: 10.1016/j.bbrep.2025.102097. eCollection 2025 Sep.
CA 15-3 and CA 27-29 are widely used serum biomarkers for breast cancer with limited utility due to low sensitivity in early-stage disease. This study details the discovery of BF-09, a new breast cancer marker with potential for wider application.
Antibodies were screened against tumor biopsy extracts (n = 115) relative to non-cancer (n = 190) specimens using an immunoscreening array. Western blot was done to determine the molecular weight of the antibody target and test secretion in cell culture medium. A sandwich ELISA was developed and tested on mouse xenograft serum to confirm in vivo secretion. A preliminary set of cancer (n = 14) and non-cancer (n = 13) human serum samples was similarly tested to confirm the biomarker could be measured in human blood. HuProt Human Protein Array was used for target protein identification.
BF-09 antibody was selected for further study because its protein target was elevated in both early and late-stage cancer extracts (p < 0.05). On Western blot, BF-09 antibody reacted against a protein band of around 15 kDa. ELISA results confirmed that the biomarker was secreted in both mouse and human serum in quantifiable amounts. BF-09 protein had higher median concentration in serum of cancer patients (p < 0.0001) compared to non-cancer patients. Phage display and HuProt array revealed the protein target as SAGA-associated factor 29.
Due to its elevated presence in early-stage tumor tissue and its measurable secretion into serum, BF-09 is a promising breast cancer marker for further study.
CA 15-3和CA 27-29是广泛应用于乳腺癌的血清生物标志物,但由于在疾病早期敏感性较低,其应用受到限制。本研究详细介绍了一种具有更广泛应用潜力的新型乳腺癌标志物BF-09的发现。
使用免疫筛选阵列,针对肿瘤活检提取物(n = 115)与非癌标本(n = 190)筛选抗体。进行蛋白质印迹法以确定抗体靶点的分子量,并检测其在细胞培养基中的分泌情况。开发了一种夹心酶联免疫吸附测定法(ELISA),并在小鼠异种移植血清上进行测试以确认体内分泌情况。同样对一组初步的癌症患者(n = 14)和非癌症患者(n = 13)的血清样本进行测试,以确认该生物标志物可在人血液中检测到。使用HuProt人类蛋白质阵列进行靶蛋白鉴定。
选择BF-09抗体进行进一步研究,因为其蛋白靶点在早期和晚期癌症提取物中均升高(p < 0.05)。在蛋白质印迹法中,BF-09抗体与一条约15 kDa的蛋白带发生反应。ELISA结果证实,该生物标志物在小鼠和人血清中均有可定量分泌。与非癌症患者相比,癌症患者血清中BF-09蛋白的中位浓度更高(p < 0.0001)。噬菌体展示和HuProt阵列显示,该蛋白靶点为SAGA相关因子29。
由于BF-09在早期肿瘤组织中含量升高且可分泌到血清中进行检测,它是一种有前景的乳腺癌标志物,值得进一步研究。