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Affinity chromatography of human IgG with octapeptide ligands identified from eleven peptide-ligand candidates

Paper ID Volume ID Publish Year Pages File Format Full-Text
2771 131 2016 8 PDF Available
Title
Affinity chromatography of human IgG with octapeptide ligands identified from eleven peptide-ligand candidates
Abstract

•Eleven peptide-ligand candidates for human IgG were evaluated in a wide pH range.•FYCHWQDE and FYCHNQDE were identified as high affinity ligands for human IgG.•The binding sites on hIgG for the two octapeptide ligands were similar to Protein A.

We have reported earlier on the characterization of four peptides from a library of 15 peptide ligand candidates for human IgG (hIgG) obtained by a biomimetic design strategy and identified three high-affinity octapeptides. In this work, the left 11 peptides were evaluated and we found two more octapeptides (FYCHWQDE and FYCHNQDE) that showed high affinity for hIgG. The binding pH ranges for the two ligands were different, but the optimum pH values were the same for each other (pH 6.0). Both the ligands showed high specificity and bound hIgG mainly by electrostatic interactions. Ligand binding competition experiments revealed that the binding sites on hIgG for the two octapeptides were similar to those for Protein A. Finally, hIgG was purified from human serum with high purities and recovery yields with the two peptide affinity columns. Thus, among the 15 peptide candidates, a total of five octapeptides were identified as high-affinity ligands of hIgG. The five ligands were all derived from the same peptide model FYxHxxxE (where x denotes any amino acid) and contained four common hot spots F132, Y133, H137, and E143 of the affinity motif of Protein A. Analyses and evaluation of the peptide library would help deepen our understanding of the affinity binding of Protein A to IgG, and promote application of the biomimetic strategy in the design of affinity ligands for different proteins.

Keywords
Protein; Affinity; Adsorption; Chromatography; Purification; Human immunoglobulin G
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Affinity chromatography of human IgG with octapeptide ligands identified from eleven peptide-ligand candidates
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 107, 15 March 2016, Pages 18–25
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us