Improved Non-chromatographic Purification of a Recombinant Protein by Cationic Elastin-like Polypeptides
| Autor: | Dong Woo Lim, Ashutosh Chilkoti, and J. Andrew MacKay, Kimberly Trabbic-Carlson |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
animal structures
Polymers and Plastics Recombinant Fusion Proteins Molecular Sequence Data Lysine Bioengineering Sodium Chloride Biology medicine.disease_cause Article law.invention Biomaterials Residue (chemistry) Thioredoxins Peptide Library law Cations Escherichia coli Materials Chemistry medicine Amino Acid Sequence Peptide sequence Chromatography Base Sequence Molecular mass Cationic polymerization Elastin Biochemistry Recombinant DNA Thioredoxin Peptides |
| Zdroj: | Biomacromolecules. 8:1417-1424 |
| ISSN: | 1526-4602 1525-7797 |
| DOI: | 10.1021/bm060849t |
| Popis: | This paper reports an improvement in the purification of thioredoxin (Trx) expressed from E. coli by inverse transition cycling (ITC) using cationic elastin-like polypeptides (ELPs). Two ELP libraries having 2% and 5% lysine residues and molecular weights ranging from 4 to 61.1 kDa showed greater salt sensitivity in their inverse transition behavior than purely aliphatic ELPs. Expression yield of Trx-ELP fusions was an unpredictable function of guest residue composition, but reducing the molecular weight of the ELP tag generally increased Trx yield. A cationic 4.3 kDa ELP is the shortest ELP used to purify any protein by ITC to date. A 15.9 kDa ELP with a guest residue composition of K:V:F of 1:7:1 was found to be the optimal cationic tag to purify Trx, as it provided 50% greater Trx yield and only required one-fifth the added NaCl for purification of Trx as compared to previously used aliphatic ELP tags. |
| Databáze: | OpenAIRE |
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