Journal
BIOCONJUGATE CHEMISTRY
Volume 27, Issue 7, Pages 1569-1573Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.bioconjchem.6b00170
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Funding
- NSFC [31270871, 21502103]
- MOEC [2012Z02293]
- Chinese Ministry of Science and Technology [2014CBA02005, 2014AA020907]
- Global Science Alliance Pro-Gram of Thermo-Fisher Scientific
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Development of a peptide-based affinity matrix and detection reagent is important for biomedical research and the biopharmaceutical industry. In the present work, we designed and synthesized an immunoglobin G (IgG)-binding peptide ligand, Fc-III-4C. Fc-III-4C is composed of 15 residues, and the 4 cysteine residues form 2 disulfide bonds to generate a double cyclic structure. The binding affinity of the Fc-III-4C peptide toward human IgG was determined to be 2.45 nIvI (K-d), which is higher than that of IgG with Protein A/G (Pro-A/G). Importantly, the Fc-III-4C peptide displayed high affinity to various IgGs from different species. Fc-III-4C immobilized agarose beads exhibited high stability and reusability when compared with that of the Pro-A/G-immobilized beads. The conjugate of Fc-III-4C with FITC was demonstrated to be suitable for immunofluorescence detection of proteins expressed in cells. These results demonstrate that the Fc-III-4C peptide is a useful affinity ligand for antibody purification and as a protein detection reagent.
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