论文部分内容阅读
转化生长因子-β_1(TGF-β_1)是转化生长因子家族中的一个重要成员,具有多种生物学功能,特别是调节细胞的生长和分化,参与细胞外基质合成,创伤愈合和胚胎形态发生等生物学过程。近年来,发展了一些抗TGF-β_1多克隆抗体和个别重组人TGF-β_1的单克隆抗体,对研究各种细胞的TGF-β_1的合成,定位和其它生物学效应发挥了作用。本文报道了抗人血小板TGF-β_1单克隆抗体TB 21的制备及其主要生物学特性,尤其是它对TGF-β_1免疫识别的专一性,以及调变对TGF-β_1敏感细胞的生长和增殖抑制的活性。结果表明,TB_(21)单克隆抗体为IgG_1亚型;对TGF-β_1有较高亲和力,亲和常数(Kaff)为1.47×10~8M~(-1);Westernblot 显示TB_(21)抗体能专一地结合25 Kd 成熟型分子和其12.5 Kd 的单体分子,对CCL/64细胞生长抑制和对NRK-49 F 成纤维细胞软琼脂集落形成的鉴定都证明TB21单抗对TGF-β_1的生物学效应有专一性的中和作用,可用于TGF-β_1的有关生物学研究。
Transforming growth factor-β_1 (TGF-β_1) is an important member of the transforming growth factor family and possesses a variety of biological functions, especially regulating cell growth and differentiation, participating in extracellular matrix synthesis, wound healing and embryogenesis Biological process. In recent years, some anti-TGF-β 1 polyclonal antibodies and several recombinant human TGF-β 1 monoclonal antibodies have been developed to study the synthesis, localization and other biological effects of TGF-β 1 in various cells. This article reports the preparation and the main biological characteristics of anti-human platelet TGF-β 1 monoclonal antibody TB 21, especially its specificity for immune recognition of TGF-β 1 and the modulation of the growth and proliferation of TGF-β 1-sensitive cells Inhibitory activity. The results showed that the TB_ (21) monoclonal antibody was IgG_1 subtype, had high affinity to TGF-β_1, and the affinity constant (Kaff) was 1.47 × 10 -8 M -1. Western blot showed that TB_ (21) Specific inhibition of both CCL / 64 cell growth and soft agar colony formation on NRK-49 F fibroblasts by binding both the 25 Kd mature molecule and its 12.5 Kd monomeric molecule demonstrated that TB21 mAb alone Biological effects of specific neutralization, can be used for TGF-β_1 related biological research.