PMID- 36002416 OWN - NLM STAT- MEDLINE DCOM- 20220826 LR - 20220826 IS - 1872-2075 (Electronic) IS - 1000-3061 (Linking) VI - 38 IP - 8 DP - 2022 Aug 25 TI - [A method for immortalizing swine monoclonal B cells secreting anti-PRRSV antibodies]. PG - 2872-2882 LID - 10.13345/j.cjb.210856 [doi] AB - Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established. FAU - Wang, Jian AU - Wang J AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Zhang, Jing AU - Zhang J AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Li, Kun AU - Li K AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Sun, Pu AU - Sun P AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Li, Guoxiu AU - Li G AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Li, Jiaoyang AU - Li J AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Cao, Yimei AU - Cao Y AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Zhao, Zhixun AU - Zhao Z AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Yuan, Hong AU - Yuan H AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Fu, Yuanfang AU - Fu Y AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Li, Pinghua AU - Li P AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Li, Dong AU - Li D AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Liu, Zaixin AU - Liu Z AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. FAU - Lu, Zengjun AU - Lu Z AD - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China. AD - College of Veterinary Medicine, Lanzhou University, Lanzhou 730046, Gansu, China. LA - chi PT - Journal Article PL - China TA - Sheng Wu Gong Cheng Xue Bao JT - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JID - 9426463 RN - 0 (Antibodies, Monoclonal) RN - 0 (Antibodies, Neutralizing) RN - 0 (Antibodies, Viral) SB - IM MH - Animals MH - Antibodies, Monoclonal MH - Antibodies, Neutralizing MH - Antibodies, Viral MH - Humans MH - *Porcine Reproductive and Respiratory Syndrome/prevention & control MH - *Porcine respiratory and reproductive syndrome virus/genetics MH - Swine OTO - NOTNLM OT - B cell immortalization OT - PRRSV OT - magnetic bead method OT - monoclonal antibody EDAT- 2022/08/25 06:00 MHDA- 2022/08/27 06:00 CRDT- 2022/08/24 22:42 PHST- 2022/08/24 22:42 [entrez] PHST- 2022/08/25 06:00 [pubmed] PHST- 2022/08/27 06:00 [medline] AID - 10.13345/j.cjb.210856 [doi] PST - ppublish SO - Sheng Wu Gong Cheng Xue Bao. 2022 Aug 25;38(8):2872-2882. doi: 10.13345/j.cjb.210856.