Fishwild D M, O'Donnell S L, Bengoechea T, Hudson D V, Harding F, Bernhard S L, Jones D, Kay R M, Higgins K M, Schramm S R, Lonberg N
Department of Hybridoma Development, GenPharm International, Mountain View, CA 94043, USA.
Nat Biotechnol. 1996 Jul;14(7):845-51. doi: 10.1038/nbt0796-845.
Human immunoglobulin transgenic mice provide a method of obtaining human monoclonal antibodies (Mabs) using conventional hybridoma technology. We describe a novel strain of human immunoglobulin transgenic mice and the use of this strain to generate multiple high-avidity human sequence IgG kappa Mabs directed against a human antigen. The light chain transgene is derived in part from a yeast artificial chromosome clone that includes nearly half of the germline human V kappa region. In addition, the heavy-chain transgene encodes both human mu and human gamma 1 constant regions, the latter of which is expressed via intratransgene class switching. We have used these animals to isolate human IgG kappa Mabs that are specific for the human T-cell marker CD4, have high binding avidities, and are immunosuppressive in vitro. The human Mab-secreting hybridomas display properties similar to those of wild-type mice including stability, growth, and secretion levels. Mabs with four distinct specificities were derived from a single transgenic mouse, consistent with an extensive diversity in the primary repertoire encoded by the transgenes.
人免疫球蛋白转基因小鼠提供了一种利用传统杂交瘤技术获得人单克隆抗体（Mab）的方法。我们描述了一种新型的人免疫球蛋白转基因小鼠品系，以及利用该品系产生多种针对人抗原的高亲和力人源序列IgG κ单克隆抗体的方法。轻链转基因部分来源于一个酵母人工染色体克隆，该克隆包含近一半的种系人Vκ区域。此外，重链转基因编码人μ链和人γ1恒定区，后者通过转基因内类别转换表达。我们利用这些动物分离出了对人T细胞标志物CD4具有特异性、具有高结合亲和力且在体外具有免疫抑制作用的人IgG κ单克隆抗体。分泌人单克隆抗体的杂交瘤表现出与野生型小鼠相似的特性，包括稳定性、生长和分泌水平。具有四种不同特异性的单克隆抗体源自一只转基因小鼠，这与转基因编码的初级库中的广泛多样性一致。
