稳定、高亲和力人源抗体的高效制备方法研究- - 抗TNF-α骆驼化人源新型基因工程抗体HcAb的制备及评价

Antibodies are enjoying wide spread use in research, diagnostic and treatment. However, antibody engineering is facing with strict problems in stability, affinity and immunogenicity at present. Moreover, the human antibodies research of our nation developed rapidly in the last 10 years, and the related technologies and estimating systems need to be improved and optimized..To develop efficient methods of preparing stable human antibodies with high affinity, and to complete the related antibody estimating technology, this study will be conducted to prepare human camelized heavy chain antibodies (HcAbs) using eukaryotic expression system. Based on the last grant supported by NSFC, the produced single-domain antibodies (sdAbs) which are specific for sTNF-α will be act as original antibodies to produce new-born human camelized HcAbs. The specific anti-TNF-α HcAbs with high affinity will be screened by the tmTNF-α expressed on cellular surface as targeting antigen. Then the related methods and technology will be developed to estimate the characteristics and the neutralizing abilities of HcAbs in vivo and in vitro. So this study will pave the way for effective applications of anti-TNF-α antibodies in patients. And it also will propel the preparation and estimation of other human engineering antibodies in our country.

近年来，抗体已广泛应用于科学研究、疾病诊断及防治等领域，但越来越多的应用对抗体的稳定性、亲和力及其人源化程度提出了更高的要求。目前我国的人源抗体研究处于快速发展阶段，其高效制备及评价的相关技术平台亟需构建和完善，工程抗体研究正面临着严峻挑战。.为建立稳定、高亲和力人源抗体的高效制备方法及评价技术体系，本项目拟在前期研究基础上，以获得的稳定、高亲和力针对sTNF-α的骆驼化人源sdAb为母本抗体，构建骆驼化人源新型基因工程抗体HcAb，并实现其在哺乳动物细胞中的高效表达；以真核细胞表面表达的tmTNF-α作为靶标抗原，进一步筛选鉴定具有临床应用价值的高亲和力特异抗TNF-α人源抗体；基于转基因动物模型等方法，完成该新型抗体的生物学特性和中和活性评价。从而为制备有临床应用价值的抗TNF-α人源抗体奠定基础，并为其他人源工程抗体的高效制备及评价提供理论依据和技术支持，进而推动我国的人源抗体研究。

为建立稳定、高亲和力人源抗体的高效制备方法及评价技术体系，本项目在前期研究基础上，以获得的稳定、高亲和力针对sTNF-α的骆驼化人源sdAb为母本抗体，构建了骆驼化人源新型基因工程抗体HcAb，并实现了其在哺乳动物细胞中的高效表达；以真核细胞表面表达的tmTNF-α作为靶标抗原，进一步筛选鉴定具有临床应用价值的高亲和力特异抗TNF-α人源抗体；并完成了该新型抗体的生物学特性和中和活性评价。本研究为制备有临床应用价值的抗TNF-α人源抗体奠定了基础，并为其他人源工程抗体的高效制备及评价提供理论依据和技术支持。