Hansaem Lee
Korea Centers for Disease Control & Prevention, Republic of Korea
Title: Comparison of three human B cell isolation tools for the development of the antibodies neutralizing emerging infectious viruses
Biography
Biography: Hansaem Lee
Abstract
Isolating human B cell cells is inevitable to investigate the development of protective antibodies, as vaccine or therapeutics candidates, against emerging infectious viruses. Single B cell sorting is a powerful tool for define ant-viral B cells. Thus, the acquisition of abundant B cells is essential to synthesize the broad-ranged antibody cDNAs for the screening of the broadly neutralizing monoclonal antibodies against the infectious viruses. In this study, we test and compare the human B cell isolation rates among the three human B cell isolation tools, a fluorescence-activated cell sorting (FACS), EasySep™ human B cell enrichment kit (STEMCELL Technologies), and human CD19 MciroBeads (MACS Miltenyi Biotec). We found that the method using FACS was more suitable to obtain more human B cells than the other methods. In addition, using a CMV tetramer probe or MERS-CoV spike protein probes, the antibody genes against CMV and MERS-CoV were amplified by heavy chain and light chain RT-PCR and nested PCR successfully. These genes were also analyzed by IMGT, resulting that the antibody sequences were proper IgG genes. The results demonstrate that human B cell isolation using FACS are a reliable tool for assisting in the rapid development of prophylactic or therapeutic antibodies neutralizing new emerging infectious diseases.