Kilpatrick K E, Cutler T, Whitehorn E, Drape R J, Macklin M D, Witherspoon S M, Singer S, Hutchins J T
Department of Molecular Sciences, Glaxo Wellcome, Research Triangle Park, NC 27709, USA.
Hybridoma. 1998 Dec;17(6):569-76. doi: 10.1089/hyb.1998.17.569.
Class-switched, affinity-matured murine monoclonal antibody (MAb)-producing cell lines were generated against the Flt-3 receptor in less than 4 weeks following polynucleotide immunizations, used in conjunction with repetitive immunizations, multiple sites (RIMMS). Plasmid DNA encoding Flt-3/Fc was coated onto gold particles, which were subsequently propelled into the epidermis of mice using biolistic particle bombardment using the Accell gene gun. Pools of immune peripheral lymph node cells were somatically fused 13 days after the onset of delivery of DNA encoding the target antigen. To determine if early responses could be augmented, DNA-encoding murine GM-CSF was delivered 3 days prior to the Flt-3/Fc DNA immunizations. The data presented demonstrates the successful identification and characterization of class-switched, affinity-matured MAbs that bind to the Flt-3 receptor. When compared to conventional methodologies or intramuscular targeted DNA-based immunization for the generation of MAbs, use of the gene gun in conjunction with RIMMS allows for a more rapid production of affinity-matured MAb-producing cell lines.
在多核苷酸免疫接种后不到4周内，结合重复免疫多部位接种法（RIMMS），产生了针对Flt-3受体的类别转换、亲和力成熟的小鼠单克隆抗体（MAb）产生细胞系。将编码Flt-3/Fc的质粒DNA包被在金颗粒上，随后使用Accell基因枪通过生物弹道粒子轰击将其推进小鼠表皮。在开始递送编码靶抗原的DNA 13天后，将免疫外周淋巴结细胞池进行体细胞融合。为了确定早期反应是否可以增强，在Flt-3/Fc DNA免疫接种前3天递送编码小鼠GM-CSF的DNA。所呈现的数据表明成功鉴定和表征了与Flt-3受体结合的类别转换、亲和力成熟的单克隆抗体。与用于产生单克隆抗体的传统方法或基于肌肉靶向DNA的免疫接种相比，使用基因枪结合RIMMS能够更快速地产生亲和力成熟的单克隆抗体产生细胞系。
