Apparent heterogeneity in the pIII-peptide fusion protein in single-phage clones isolated from peptide libraries
| Autor: | Bo Mattiasson, Igor Yu. Galaev, Wim Noppe, Hans Deckmyn |
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| Rok vydání: | 2011 |
| Předmět: |
Recombinant Fusion Proteins
Antibody Affinity Bioengineering Peptide Biology Protein Engineering Biochemistry Chromatography Affinity Affinity chromatography Peptide Library Escherichia coli Humans Bacteriophages Avidity Peptide library Molecular Biology chemistry.chemical_classification Protein engineering Flow Cytometry Ligand (biochemistry) Molecular biology Affinities Fusion protein Lactoferrin chemistry Oligopeptides Cryogels Biotechnology |
| Zdroj: | Protein Engineering Design and Selection. 24:721-726 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/gzr033 |
| Popis: | Ligand homogeneity is an important issue in affinity chromatography. Using phages expressing peptides on the pIII protein, a heterogeneity in the binding of monoclonal phages was observed during affinity chromatography on supermacroporous cryogels. Fractions with different apparent binding affinities could be separated by stepwise elution. When these different fractions were re-applied, the respective differences in affinity were retained. However, when phage fractions with different apparent affinities were first amplified, an offspring was generated with again variable affinities. As the sequence of the peptide insert was the same, the heterogeneity must be ascribed to differences in avidity and although no direct evidence could be generated, we hypothesize that this is possibly due to phages displaying different numbers of the same peptide as a consequence of either proteolytic or packaging events during the amplification step in Escherichia coli. |
| Databáze: | OpenAIRE |
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