Engineering a cell-penetrating hyperstable antibody scFv(Ras) – An extraordinary approach to cancer therapeutics

Abstract

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).

Antibody variable region engineering for improving cancer immunotherapy

“…The solid tumor is relatively inaccessible to whole antibodies because of several biological barriers [ 21 ]. In contrast, scFv and nanobodies have superior tissue penetration properties compared with mAbs: they demonstrate a higher perfusion rate across endothelial and epithelial barriers and diffuse faster through the tumor stroma because of their smaller size [ 22 , 23 , 24 , 25 ]. Nevertheless, the small size of scFv and nanobodies also has its drawbacks.…”

Ecological Environment and Pollution Control under the Influence of Modern Biotechnology Development

“…The fourth is to use modern information technology to strengthen public education and publicity to improve the public's understanding of local governance. Governments at all levels should strengthen publicity and education on green environmental protection, energy conservation and emission reduction according to the actual situation and through various media platforms, such as setting up environmental protection classes in schools, so that students who have learned can understand the protection of the physical environment [7][8].…”

A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo

“…Hydrophobic peptide gH625 is a membrane-perturbing domain, which interacts with biological membranes and is able to transport different cargoes into the cytoplasm [ 18 ]. Due to CPPs’ non-specific mediation, coupling monoclonal antibodies to CPP-functionalized carriers has emerged as an effective strategy for enhancing targeted delivery, with significant recent advancements [ 19 , 20 , 21 ]. Lacasse et al were the first to report a cholic-acid-linked conjugate, combining a nuclear localization signal sequence with trastuzumab-emtansine (T-DM1).…”