Maestre Lorena, Fontan Lorena, Martinez-Climent Jose Angel, Garcia Jose Francisco, Cigudosa Juan Cruz, Roncador Giovanna
Monoclonal Antibodies Unit, Biotechnology Programme, Spanish National Cancer Centre (CNIO), Madrid, Spain.
Hybridoma (Larchmt). 2007 Apr;26(2):86-91. doi: 10.1089/hyb.2006.044.
Genetic immunization (GI), which is primarily used for vaccine purposes, is a method for producing antibodies to a protein by delivering the gene encoding the protein as a eukaryotic expression vector instead of the protein itself. The mucosa-associated lymphoid tissue lymphoma translocation gene 1 (MALT1) is one of the most likely candidates for involvement in pathogenesis of MALT lymphoma and probably of multiple myelomas. In the present work we describe the production and characterization of a mouse monoclonal antibody (mAb) directed against MALT1 and the study of MALT1 protein expression in a large series of lymphomas and myeloma cell lines. The full-length coding sequence of human MALT1 was inserted into pcDNA3 vector and delivered into mouse skin using a helium gene gun. Six new mAbs against the MALT1 molecule were produced. In order to characterize and confirm the specificity of these mAbs, Western blot (WB) and immunoprecipitation (IP) analyses were performed. A new anti-MALT1 mAb was selected and tested in a large series of cell lines. These results confirm that GI is a reliable and effective alternative method for production of mAbs, allowing accurate and sensitive detection and screening of proteins by WB.
基因免疫(GI)主要用于疫苗目的,它是一种通过将编码蛋白质的基因作为真核表达载体而非蛋白质本身来产生针对该蛋白质的抗体的方法。黏膜相关淋巴组织淋巴瘤易位基因1(MALT1)是最有可能参与MALT淋巴瘤发病机制的候选基因之一,可能也参与多发性骨髓瘤的发病机制。在本研究中,我们描述了一种针对MALT1的小鼠单克隆抗体(mAb)的制备和特性鉴定,以及对一系列淋巴瘤和骨髓瘤细胞系中MALT1蛋白表达的研究。将人MALT1的全长编码序列插入pcDNA3载体,并使用氦气基因枪将其导入小鼠皮肤。制备了六种针对MALT1分子的新mAb。为了鉴定和确认这些mAb的特异性,进行了蛋白质印迹(WB)和免疫沉淀(IP)分析。选择了一种新的抗MALT1 mAb并在一系列细胞系中进行了测试。这些结果证实,基因免疫是一种可靠且有效的制备单克隆抗体的替代方法,能够通过蛋白质印迹准确、灵敏地检测和筛选蛋白质。