Selection for a periplasmic factor improving phage display and functional periplasmic expression
| Autor: | Hendrick Bothmann, Andreas Plückthun |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Phage display
Recombinant Fusion Proteins Two-hybrid screening Phagemid Biomedical Engineering chemical and pharmacologic phenomena Bioengineering Biology medicine.disease_cause Coliphages Applied Microbiology and Biotechnology law.invention Bacteriophage Bacterial Proteins law Escherichia coli medicine Cloning Molecular Immunoglobulin Fragments Escherichia coli Proteins Periplasmic space respiratory system biology.organism_classification Fusion protein Molecular biology DNA-Binding Proteins Biochemistry Genes Bacterial Periplasm Recombinant DNA Molecular Medicine Molecular Chaperones Biotechnology |
| Zdroj: | Nature Biotechnology. 16:376-380 |
| ISSN: | 1546-1696 1087-0156 |
| Popis: | The efficiency of both phage display in Escherichia coli and periplasmic expression of recombinant proteins may be limited by the same periplasmic folding steps. To search for E. coli factors that improve the efficiency of both procedures, a library of E. coli proteins was coexpressed in a phagemid vector that contained a poorly folding single-chain Fv antibody (scFv) fragment fused to g3p. We enriched, by panning for antigen binding, those phagemids in which the amount of displayed scFv is highest. We thus identified the periplasmic protein Skp/OmpH/HlpA as improving phage display of a wide range of scFv fragments. This occurs as a result of an increase in the amount of hybrid protein displayed on the phage. Coexpression of skp also increases the functional yield of scFv fragments when expressed by secretion to the periplasm. |
| Databáze: | OpenAIRE |
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