【 摘 要 】
A fully functional cysteine-free derivative of the intrinsically stable anti-HER2 scFv fragment hu4D5–8 was generated by replacing the disulfide forming cysteine residues in V H and V L with the amino acid combination valine-alanine in both domains. The antigen binding properties, determined by ELISA and BIAcore measurements, were not affected by removal of the disulfide bonds. The thermodynamic stability of the disulfide-containing scFv of 8.1 kcal/mol is decreased upon complete reduction of both disulfides to 2.7 kcal/mol, while that of the valine-alanine variant is somewhat higher (about 3.8 kcal/mol). Our results suggest that, in principle, a disulfide-free fully functional derivative of any scFv can be obtained, as long as the corresponding disulfide-containing scFv has a high enough thermodynamic stability.
【 授权许可】
Unknown
【 预 览 】
Files | Size | Format | View |
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RO201912020305949ZK.pdf | 425KB | download |