التفاصيل البيبلوغرافية
| العنوان: |
Purification of a Novel Anti-VEGFR2 Single Chain Antibody Fragment and Evaluation of Binding Affinity by Surface Plasmon Resonance. |
| المؤلفون: |
Kordi, Shirafkan, Rahmati-Yamchi, Mohammad, Asghari Vostakolaei, Mehdi, Barzegari, Abolfazl, Abdolalizadeh, Jalal |
| المصدر: |
Advanced Pharmaceutical Bulletin; 2019, Vol. 9 Issue 1, p64-69, 6p |
| مصطلحات موضوعية: |
POLYACRYLAMIDE gel electrophoresis, SURFACE plasmon resonance, VASCULAR endothelial growth factor receptors, SODIUM dodecyl sulfate |
| مستخلص: |
Purpose: The single-chain variable fragment (scFv) domain of antibodies is now considered as one of the therapeutic tools that can be produced by phage display technology (PDT). Antibody purification is one of the most important steps in antibodies production. The aim of study was purification and characterization of anti-VEGFR2 scFv antibody fragments. Methods: After the coating of vascular endothelial growth factor receptor 2 (VEGFR2) peptide in ELISA microplates, the phage display library of Tomlinson was used for antibody isolation. The targeted scFv was purified by chromatography using a zeolite-based column. The purity and functional assessment of purified scFv were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting techniques, respectively. Affinity binding was evaluated by surface plasmon resonance (SPR). Results: The desired scFv was selected after four stages of biopanning. SDS-PAGE analysis showed a 28 kDa scFv with high purity (>90%). The western bloting analysis confirmed the binding of produced scFv antibody to the desired peptide. The affinity binding of scFv antibody analyzed by SPR was about 60 μM. Conclusion: In this study, the novel scFv antibody against VEGFR2 peptide was purified by chromatography column containing zeolite. Based on our findings the produced antibody may be applied for diagnosis or targeting of VEGFR2 in antibody-based therapy strategies. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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