Isolation of a Novel Anti-KDR3 Single-chain Variable Fragment Antibody from a Phage Display Library
| Autor: | Shirafkan Kordi, Mohammad Rahmati-Yamchi, Mehdi Asghari Vostakolaei, Ali Etemadie, Abolfazl Barzegari, Jalal Abdolalizadeh |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: | |
| Zdroj: | Iranian Journal of Allergy, Asthma and Immunology, Vol 18, Iss 3 (2019) |
| Druh dokumentu: | article |
| ISSN: | 1735-1502 1735-5249 |
| DOI: | 10.18502/ijaai.v18i3.1122 |
| Popis: | Vascular endothelial growth factor receptor 2 (VEGFR-2) is known as one of the important antigens playing a vital role in angiogenesis. In this study, phage display technology (PDT) was used to produce a single-chain variable fragment (scFv) antibody against a region of the domain 3 in VEGFR-2 called kinase insert domain receptor 3 (KDR3). After designing the KDR3 peptide and biopanning, a colony was chosen for scFv antibody expression. Following expression and purification; western blotting, dot blotting and immunofluorescence (IF) were used to evaluate the antibody function. Surface plasmon resonance (SPR) was also employed to measure affinity of produced antibody. Once a colony was selected and transferred to the expression host, the scFv antibody was expressed in the expected range of 28 kDa. Using a designed chromatography column, antibody purification was found to be about 95%. In this study, a novel scFv with the capability of binding to KDR3 was isolated and purified and its intracellular function was investigated and verified. |
| Databáze: | Directory of Open Access Journals |
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