Use of phage display methods to identify heptapeptide sequences for use as affinity purification 'tags' with novel chelating ligands in immobilized metal ion affinity chromatography
| Autor: | Jane Therese Mooney, Milton Thomas William Hearn, Dale Patricia Fredericks |
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| Rok vydání: | 2010 |
| Předmět: |
Phage display
Recombinant Fusion Proteins Peptide Biochemistry Chromatography Affinity Analytical Chemistry law.invention Affinity chromatography law Heterocyclic Compounds Nickel Peptide Library Chelation Histidine Amino Acids Peptide library chemistry.chemical_classification Chromatography Chemistry Organic Chemistry General Medicine Chromatography Ion Exchange Amino acid Recombinant DNA Electrophoresis Polyacrylamide Gel Peptides Protein Binding |
| Zdroj: | Journal of chromatography. A. 1218(1) |
| ISSN: | 1873-3778 |
| Popis: | This study describes the screening of a peptide phage display library for amino acid sequences that bind with different affinities to a novel class of chelating ligands complexed with Ni²+ ions. These chelating ligands are based on the 1,4,7-triazacyclononane (TACN) structure and have been chosen to allow enhanced efficiency in protein capture and decreased propensity for metal ion leakage in the immobilized metal ion affinity chromatographic (IMAC) purification of recombinant proteins. Utilising high stringency screening conditions, various peptide sequences containing multiple histidine, tryptophan, and/or tyrosine residues were identified amongst the different phage peptide sequences isolated. The structures, and particularly the conserved locations of these key amino acid residues within the selected heptapeptides, form a basis to design specific peptide tags for use with these novel TACN ligands as a new mode of IMAC purification of recombinant proteins. |
| Databáze: | OpenAIRE |
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