An efficient method for purification of human T-cell growth factor
| Autor: | M. Nabholz, M Cianfriglia, B. Chapuis, Oreste Acuto, M. Colombatti |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
medicine.medical_treatment
Immunology Size-exclusion chromatography Polyethylene glycol Biology Cell Line chemistry.chemical_compound Mice PEG ratio medicine Immunology and Allergy Animals Chemical Precipitation Humans Ammonium Phytohaemagglutinin Chromatography Growth factor Chromatography Agarose Culture Media chemistry Cell culture Chloramine-T biology.protein Chromatography Gel Interleukin-2 Electrophoresis Polyacrylamide Gel Dialysis Spleen T-Lymphocytes Cytotoxic |
| Popis: | A 1000-fold purification of human T-cell growth factor (TCGF) was achieved starting from supernatants of human spleen cells stimulated with phytohaemagglutinin (PHA) in culture medium containing 0.5% serum. The purification scheme involved precipitation with ammonium sulphate, gel filtration and blue-Sepharose chromatography. The use of polyethylene glycol 6000 (PEG 6000) was critical during the chromatographic steps in order to obtain high final recoveries or activity (40-50%). Purified preparations of TCGF labelled with 125I by the chloramine T method revealed that the activity co-migrated with 2 molecular species of 14,000-17,000 daltons in SDS-PAGE under non-reducing conditions. |
| Databáze: | OpenAIRE |
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