Production of Leptin inEscherichia coli:A Comparison of Methods
| Autor: | Jeffrey P Varnerin, T. Smith, Aurawan Vongs, Patricia J. Hey, M. Chou, Joseph J. King, Theodore N. Mellin, Easter G. Frazier, Michael R. Tota, D. E. Macintyre, Beth Junker, Beth Ann Murphy, Charles Rosenblum, Christian N. Nunes, B. W. Burgess, L. H. T. Van Der Ploeg |
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| Rok vydání: | 1998 |
| Předmět: |
Leptin
Protein Folding medicine.medical_specialty Recombinant Fusion Proteins medicine.medical_treatment Genetic Vectors Radioimmunoassay Mice Obese Inclusion bodies Cell Line Mice In vivo Internal medicine Weight Loss Escherichia coli medicine Animals Humans Histidine Inclusion Bodies Leptin receptor Chemistry Insulin digestive oral and skin physiology Proteins Mice Mutant Strains Endotoxins Diafiltration Endocrinology Biochemistry Protein Biosynthesis Biological Assay Peptides Filtration hormones hormone substitutes and hormone antagonists Ex vivo Biotechnology |
| Zdroj: | Protein Expression and Purification. 14:335-342 |
| ISSN: | 1046-5928 |
| DOI: | 10.1006/prep.1998.0978 |
| Popis: | A procedure is described for gram-scale refolding of Escherichia coli-derived human leptin inclusion bodies. Refolding was achieved by gradually reducing denaturant using a diafiltration method. Refolded leptin is characterized by in vivo modulation of food intake, reduction in body weight, and lowering of insulin and glucose levels in ob/ob mice. In addition, refolded leptin is characterized by radioimmunoassay (RIA) and activation of the leptin receptor in a cell-based assay. For comparison we also refolded leptin by a simple dilution method and produced periplasmic derived leptin, which did not require ex vivo folding. Leptin produced by these three methods and leptin obtained from commercial sources were compared using the RIA and the cell-based assay and appeared to be of comparable quality and potency. |
| Databáze: | OpenAIRE |
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