Rothe Achim, Nathanielsz Anne, Hosse Ralf J, Oberhäuser Frank, Strandmann Elke P, Engert Andreas, Hudson Peter J, Power Barbara E
Department of Internal Medicine I, Laboratory of Immunotherapy, University Hospital Cologne, Joseph Stelzmann street 9, 50931 Cologne, Germany.
J Biotechnol. 2007 Jul 15;130(4):448-54. doi: 10.1016/j.jbiotec.2007.05.012. Epub 2007 May 24.
Engineered antibodies have become an invaluable source of biopharmaceuticals against a wide range of diseases. About 200 antibody-based biologicals have been tested in clinical trials. Single chain variable fragments of antibodies (scFvs) provide binding specificity and offer an increased ease of in vitro display selection. Here, we present the generation of a human scFv library from peripheral blood lymphocyte RNA of a patient with relapsed T-cell non-Hodgkin lymphoma (T-NHL) who experienced a rare case of "spontaneous" remission. Antibodies against human T-cell antigen CD28, a co-stimulatory protein that influences the immune response by amplification of the transcriptional effects of T-cell receptors, might have contributed to the patient's remission. The scFv library was panned against CD28 using ribosome display and further subjected to affinity maturation. Isolated scFv were assessed for binding specificity and affinity and may provide the basis for the development of novel immunotherapeutic strategies. This work demonstrates the selection of a fully human antibody fragment from a patient-derived gene pool by in vitro ribosome display technology.
工程抗体已成为对抗多种疾病的生物制药的宝贵来源。约200种基于抗体的生物制品已在临床试验中进行了测试。抗体的单链可变片段(scFv)具有结合特异性,并且在体外展示筛选方面更加简便。在此,我们展示了从一名复发的T细胞非霍奇金淋巴瘤(T-NHL)患者的外周血淋巴细胞RNA中生成人scFv文库的过程,该患者经历了罕见的“自发”缓解情况。针对人T细胞抗原CD28的抗体,CD28是一种共刺激蛋白,通过放大T细胞受体的转录效应来影响免疫反应,可能促成了患者的缓解。使用核糖体展示技术针对CD28筛选scFv文库,并进一步进行亲和力成熟。对分离出的scFv进行结合特异性和亲和力评估,可为新型免疫治疗策略的开发提供基础。这项工作展示了通过体外核糖体展示技术从患者来源的基因库中筛选出完全人源抗体片段的过程。