Development of the Double Cyclic Peptide Ligand for Antibody Purification and Protein Detection
Development of the Double Cyclic Peptide Ligand for Antibody Purification and Protein Detection
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用于抗体纯化和蛋白质检测的双环肽配体的开发
DOI：
10.1021/acs.bioconjchem.6b00170
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发表时间：
2016-07-01
影响因子：
4.7
通讯作者：
Deng, Haiteng
中科院分区：
文献类型：
--
作者：
Gong, Yiyi;Zhang, Lin;Deng, Haiteng
开发基于多肽的亲和基质和检测试剂对生物医学研究和生物制药工业具有重要意义。在本工作中，我们设计并合成了一种免疫球蛋白G(Ig G)结合肽配体Fc-III-4C。FC-III-4C由15个残基组成，4个半胱氨酸残基形成2个二硫键形成双环结构。Fc-III-4C与人免疫球蛋白的结合亲和力为2.45Nivi(K-d)，高于免疫球蛋白与蛋白A/G的结合亲和力(Pro-A/G)。重要的是，Fc-III-4C多肽与不同物种的不同免疫球蛋白具有很高的亲和力。与Pro-A/G固定化小球相比，FC-III-4C固定化小球具有更高的稳定性和可重用性。FITC与FC-III-4C的偶联物适合于细胞表达蛋白的免疫荧光检测。这些结果表明，Fc-III-4C多肽是一种用于抗体纯化和作为蛋白质检测试剂的亲和配体。
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.