Ditzel Henrik J
Institute of Medical Biology, University of Southern Denmark, Winsloewparken 25, Odense, Denmark.
Methods Mol Biol. 2009;562:37-43. doi: 10.1007/978-1-60327-302-2_3.
A method is described for affinity isolation of antigen-specific circulating B cells of interest for subsequent generation of immune antibody phage display libraries. This approach should overcome the problem of low yields of monoclonal antibodies of interest in the libraries generated from peripheral blood lymphocytes caused by the low abundance of antigen-specific B cells in the circulation. The preselection of B cells is based on the specificity of the surface Ig receptor and is accomplished using the antigen of interest conjugated to magnetic beads. This method should significantly increase the frequency of antibody phage particles of interest in the library and allow for efficient isolation monoclonal antibodies with the predefined specificity.
本文描述了一种用于亲和分离感兴趣的抗原特异性循环B细胞的方法，以便随后生成免疫抗体噬菌体展示文库。这种方法应能克服在外周血淋巴细胞产生的文库中，由于循环中抗原特异性B细胞丰度低而导致的感兴趣的单克隆抗体产量低的问题。B细胞的预选基于表面Ig受体的特异性，并使用与磁珠偶联的感兴趣的抗原完成。该方法应能显著提高文库中感兴趣的抗体噬菌体颗粒的频率，并允许高效分离具有预定义特异性的单克隆抗体。
