Baradaran Behzad, Majidi Jafar, Farajnia Safar, Barar Jaleh, Omidi Yadollah
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
Hum Antibodies. 2014;23(1-2):13-20. doi: 10.3233/HAB-140278.
Tumor growth and progression depends largely on the activity of cell membrane receptors like epidermal growth factor receptor (EGFR) that plays a pivotal role in the progression and invasion of different solid tumors. As one of the most promising approaches for targeting and therapy, monoclonal antibodies (mAbs) have widely been used in the treatment of various malignancies. However, the clinical effects of mAbs appear to be dependent upon its specificity and potency. In the current investigation, a series of mAbs were produced against human EGFR using hybridoma technology. Balb/c mice were immunized against EGFR-positive A431 cancer cells and the most immune mouse was selected for fusion and generation of anti-EGFR mAbs. Isotyping of the generated mAbs was performed by ELISA method. Of various monoclones produced, IgG1 subclass (mAb BF4) displayed specific binding to the EGFR-expressing A431 cells. Flow cytometry and immunofluorescence staining revalidated its specific reactivity with EGFR and MTT assay revealed significant growth inhibition of A431 cells treated with mAb BF4 mainly through induction of apoptosis. Based on these findings, we propose the produced anti-EGFR mAb BF4 to be exploited for diagnostic and possibly treatment of various malignancies with overexpression of EGFR.
肿瘤的生长和进展在很大程度上取决于细胞膜受体的活性，如表皮生长因子受体（EGFR），其在不同实体瘤的进展和侵袭中起关键作用。作为最有前景的靶向和治疗方法之一，单克隆抗体（mAb）已广泛用于各种恶性肿瘤的治疗。然而，单克隆抗体的临床效果似乎取决于其特异性和效力。在当前研究中，使用杂交瘤技术制备了一系列针对人EGFR的单克隆抗体。用EGFR阳性的A431癌细胞免疫Balb/c小鼠，选择免疫最强的小鼠进行融合并产生抗EGFR单克隆抗体。通过ELISA方法对产生的单克隆抗体进行亚型鉴定。在产生的各种单克隆中，IgG1亚类（单克隆抗体BF4）显示出与表达EGFR的A431细胞特异性结合。流式细胞术和免疫荧光染色再次证实了其与EGFR的特异性反应性，MTT分析显示，用单克隆抗体BF4处理的A431细胞生长受到显著抑制，主要是通过诱导细胞凋亡。基于这些发现，我们建议将产生的抗EGFR单克隆抗体BF4用于诊断以及可能用于治疗EGFR过表达的各种恶性肿瘤。
