Bae Jina, Song Yoonyee
Chadwick International, Yeonsu-gu, Incheon, 22002, South Korea.
Lansing Catholic High School, Lansing, MI, 48912, USA.
Synth Syst Biotechnol. 2021 Oct 20;6(4):343-350. doi: 10.1016/j.synbio.2021.10.002. eCollection 2021 Dec.
In the modern pharmaceutical industry, monoclonal antibodies are often used as therapeutic agents. However, they are restricted to cell surface antigens due to their inability to penetrate the outer cell membrane and maintain normal function in the reducing environment. Additionally, it can lead to cytotoxicity since it attacks cancerous cells by mimicking the human immune system. As an alternative, this study modifies the hyperstable single-chain fragment variable(scFv) antibody to eliminate cancer using its linear shape. The scFv(F8) antibody model was modified to recognize human Ras protein by altering residues in the antigen-binding site. Furthermore, a cell-penetrating peptide (CPP) was attached to the scFv(Ras) antibody model to allow entrance to the cell, creating CPP-scFv(Ras). Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE), western blotting, and the binding assay were performed to prove its effectiveness. As a result, CPP-scFv(Ras) was successfully engineered and bound to the antigen, HRas(G12V).
在现代制药行业中,单克隆抗体常被用作治疗药物。然而,由于它们无法穿透细胞膜外层并在还原环境中维持正常功能,因此仅限于作用于细胞表面抗原。此外,由于它通过模仿人体免疫系统来攻击癌细胞,可能会导致细胞毒性。作为一种替代方案,本研究对超稳定单链可变片段(scFv)抗体进行修饰,利用其线性形状来消除癌症。通过改变抗原结合位点中的残基,对scFv(F8)抗体模型进行修饰,使其能够识别人类Ras蛋白。此外,将细胞穿透肽(CPP)连接到scFv(Ras)抗体模型上,使其能够进入细胞,从而构建出CPP-scFv(Ras)。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白质印迹法和结合试验来证明其有效性。结果,成功构建了CPP-scFv(Ras),并使其与抗原HRas(G12V)结合。