The development of a rapid and efficient system to generate porcine monoclonal antibodies (mAbs) is an important step toward the discovery of critical neutralizing targets for designing rational vaccines against porcine viruses. In this study, we established a platform for producing porcine mAbs based on single cell technologies. First, we singled out an optimal donor from 507 pigs based on serum antibody neutralizing activity against porcine reproductive and respiratory syndrome virus (PRRSV). After identifying the contribution of IgG to the neutralizing activity, single CD45R+IgG+Ag+ B cells were sorted from peripheral blood mononuclear cells (PBMCs). Single B cell RT-PCR was performed using primers designed to cover the germline repertoire of the porcine VH/VL gene segments. Paired VH/VLs were cloned into a eukaryotic expression vector and transfected into 293T cells. We demonstrate that full-length porcine mAbs were produced, and antigen-specific mAbs were obtained after further validation. The approach reported in this study can be applied to generate porcine mAbs against any given antigen and may help with the screening of neutralizing antibodies against porcine pathogens.

中文翻译：

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基于单细胞技术的猪单克隆抗体的产生。

快速高效的系统可产生猪单克隆抗体（mAbs），是朝着发现关键的中和靶标迈出的重要一步，以设计针对猪病毒的合理疫苗。在这项研究中，我们建立了一个基于单细胞技术生产猪单克隆抗体的平台。首先，我们基于针对猪繁殖与呼吸综合征病毒（PRRSV）的血清抗体中和活性，从507头猪中选出了最佳供体。在确定IgG对中和活性的贡献后，从外周血单核细胞（PBMC）分选了单个CD45R + IgG + Ag + B细胞。使用设计为覆盖猪VH / VL基因区段种系库的引物进行单B细胞RT-PCR。将配对的VH / VLs克隆到真核表达载体中，并转染到293T细胞中。我们证明了产生了全长猪单克隆抗体，并在进一步验证后获得了抗原特异性单克隆抗体。本研究报道的方法可用于产生针对任何给定抗原的猪单克隆抗体，并可能有助于筛选针对猪病原体的中和抗体。