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Preparation of anti-hCG antibody-like molecule by using a RAD peptide display system / 生物工程学报
Chinese Journal of Biotechnology ; (12): 871-879, 2019.
Article in Chinese | WPRIM | ID: wpr-771323
ABSTRACT
By using an RAD peptide display system derived from the ATPase domain of recombinase RadA of Pyrococcus furiosus, an anti-hCG antibody-like molecule was prepared by grafting an hCG-binding peptide to the RAD scaffold. After linking to sfGFP gene, a gene of hCG peptide-grafted RAD was synthesized and cloned into a bacterial expression vector (pET30a-RAD/hCGBP-sfGFP). The vector was transformed into Escherichia coli, and expression of the fusion protein was induced. After isolation and purification of the fusion protein, its binding affinity and specificity to hCG were determined by using a process of immunoabsorption followed by GFP fluorescence measurement. A comparison of hCG-binding activity with a similarly grafted single-domain antibody based on a universal scaffold was performed. The measurement of hCG-binding affinity and specificity revealed that the grafted RAD has an optimally high binding affinity and specificity to hCG, which are better than the grafted single-domain antibody. Moreover, the affinity and specificity of grafted RAD molecule are comparable to those of a commercial monoclonal antibody. In addition, the hCG-binding peptide-grafted RAD molecule has a relatively high biochemical stability, making it a good substitute for antibody with potential application.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Peptides / Recombinant Fusion Proteins / Chemistry / Escherichia coli Proteins / DNA-Binding Proteins / Escherichia coli / Genetics / Metabolism / Antibodies, Monoclonal / Antibody Specificity Limits: Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2019 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Peptides / Recombinant Fusion Proteins / Chemistry / Escherichia coli Proteins / DNA-Binding Proteins / Escherichia coli / Genetics / Metabolism / Antibodies, Monoclonal / Antibody Specificity Limits: Humans Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2019 Type: Article