Biomimetic design of affinity peptide ligands for human IgG based on protein A-IgG complex |
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| Institution: | 1. Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China;2. Synergetic Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China;1. Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;2. Dipartimento di Chimica, Materiali e Ingegneria Chimica “Giulio Natta”, Politecnico di Milano, Via Mancinelli 7, Milan 20131, Italy;1. Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA;2. Research and Development Group, Fuji Silysia Chemical Ltd., Aza Kihara, Oaza Hichiya, Hyuga-shi, Miyazaki-ken, Japan;3. Biomanufacturing Training and Education Center (BTEC), North Carolina State University, Raleigh, NC 27695, USA;4. Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA;1. Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China;2. Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin, 300072, China;1. Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou 510632, China;2. Laboratory of Analytical Pharmaceutical Chemistry, Department of Pharmaceutical Sciences, CIRM, University of Liege, CHU B36, B-4000 Liege, Belgium;3. Department of Pharmacy and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine & New Drug Research, Jinan University, Guangzhou 510632, China |
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| Abstract: | Staphylococcal protein A (SpA) has been widely used as an affinity ligand for the purification of immunoglobulin G (IgG). Based on the affinity motif of SpA, we have herein developed a biomimetic design strategy for affinity peptide ligands of IgG. First, according to the distribution of the six hot spots of the SpA affinity motif determined previously, the number of residues that should be inserted into between the hot spots was determined. Cysteine was introduced as one of the middle inserted residues of the peptide for later immobilization. Then, amino acid location was performed to identify other amino acid residues for insertion, leading to the construction of a peptide library. The library was screened by using different molecular simulation protocols, resulting in the selection of 15 peptide candidates. Thereafter, molecular dynamics simulations were performed to validate the dynamics of the affinity interactions between the candidates and IgG, and 14 of them were found to keep high affinities. Finally, the affinity and specificity of the top one ligand FYWHCLDE were exemplified by protein chromatography and IgG purification. The results indicate that the design strategy was successful and the affinity peptide ligand for IgG is promising for application in antibody purifications. |
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| Keywords: | Protein Affinity Adsorption Chromatography Purification Molecular simulation |
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