Hamid S S A, Cheah S H
Department of Physiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Hybridoma (Larchmt). 2011 Apr;30(2):137-43. doi: 10.1089/hyb.2010.0091.
Breast mucin is secreted by breast tumor cells and serves as a marker for breast cancer. Thus, antibodies against breast mucin will be valuable in the development of immunotherapy and laboratory diagnostic tests. Monoclonal antibodies (MAbs) against breast cancer-associated antigen were generated and characterized. Balb/c mice were immunized with breast cancer-associated antigen CA15-3, and subsequently splenocytes from immunized mice were fused with myeloma cells. After fusion, culture supernatants from hybridomas surviving HAT medium were screened by enzyme-linked immunosorbent assay (ELISA). A total of eight hybridomas producing MAbs against breast cancer showed significant levels of antibody activity against CA15-3. Two selected stable hybridomas were adapted into CELLine CL 350 bioreactors, and the MAbs produced were characterized for their subclass, specificity, and affinity. The MAbs were of high specificity and affinity as shown by ELISA. The MAbs produced may represent a powerful tool and are considered promising reagents for use in diagnosis and detection of early stage of the disease.
乳腺粘蛋白由乳腺肿瘤细胞分泌，可作为乳腺癌的标志物。因此，抗乳腺粘蛋白抗体在免疫治疗和实验室诊断测试的开发中具有重要价值。已制备并鉴定了针对乳腺癌相关抗原的单克隆抗体（MAb）。用乳腺癌相关抗原CA15 - 3免疫Balb/c小鼠，随后将免疫小鼠的脾细胞与骨髓瘤细胞融合。融合后，通过酶联免疫吸附测定（ELISA）筛选在HAT培养基中存活的杂交瘤细胞的培养上清液。总共8个产生抗乳腺癌单克隆抗体的杂交瘤显示出针对CA15 - 3的显著抗体活性水平。选择的两个稳定杂交瘤被接种到CELLine CL 350生物反应器中，并对产生的单克隆抗体的亚类、特异性和亲和力进行了表征。ELISA结果表明，这些单克隆抗体具有高特异性和亲和力。所产生的单克隆抗体可能是一种强大的工具，被认为是用于疾病早期诊断和检测的有前景的试剂。
