Isolation and purification of thymosin β_4 from calf thymus
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Objective:To isolate and purify thymosin β4 from calf thymus.Methods: Thymosin fraction 5 was extracted by ammonium sulfate precipitation.Thymosin β4 was purified by ion exchange chromatography,and elucidated by SDS-PAGE and HPLC.Results:The purity of thymosin β4 was 90.1%.Conclusion:Thymosin β4 was purified by ion exchange chromatography and warranted for the pilot manufacture.Keywords:
Thymosin
Ammonium sulfate precipitation
Ion chromatography
Ammonium sulfate
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Abstract Thymosin β4 (Tβ4) is a peptide composed of 43 amino acid residues with a molecular weight of 4982 and a pI of 5.1. It was originally isolated from calf thymus [Low, et al. PNAS 78, (1981)]. We have developed a rapid, efficient and reproducible method for the purification of Tβ4 from human thymus. A newly developed method was employed for the extraction of Tβ4. This extraction is simple and minimizes the possibility of proteolytic modification. This purification is based on the use of high performance reversed phase (Delta Pak C18) chromatographic columns. The HPLC retention time, RIA, IEF, and SDS-PAGE have shown that purified Tβ4 from human thymus is homogeneous and is identical to synthetic Tβ4.
Thymosin
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Thymosin
Sodium dodecyl sulfate
Fraction (chemistry)
BETA (programming language)
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Thymosin
Chromatofocusing
Homogenization
Solid phase extraction
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Protamine sulfate
Agarose
Ion chromatography
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An artificially synthesized gene of human thymosin α1(Tα1) was inserted into pBV222 and then thermo-inductively expressed as a fusion protein in a soluble form in transformed Escherichia coli strain DH5α at 42°C.The fusion protein purified through nickel affinity chromatography was cleaved with enterokinase.The cleavage mixture was returned to the nickel affinity chromatography column.Recombinant Tα1(rTα1) in the flow-through reached a purity of 94.5% as showed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and 72% by reverse phase-high performance liquid chromatography(RP-HPLC),respectively.The finally purified rTα1 exhibited the same bioactivity as chemically synthesized Tα1 in rosette tests with peripheral blood lymphocytes from cancer patients.
Enteropeptidase
Thymosin
Sodium dodecyl sulfate
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Crude synthesis thymosin α1 was isolated and purified to more than 98 5% homogeneity through ion exchange chromatography on DEAE Sephadex A50 and RepMapC18 chromatography respectively The purified thymosin α 1 appeared as a single band on the PAGE and a single peak on the HPLC The total yield was 20% The technology is simple,and can be enlarged easily
Thymosin
Ion chromatography
Sephadex
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Objective:To develop a rapid method for the purification of monoclonal B-S12 from mouse ascites with high purity, high recovery and high biological activity.Methods:The ascites was subject to precipitation with ammonium sulfate, then purified by anion exchange column chromatography. The antibody′s biological activity was tested by MTT method.Results:The method for purifying monoclonal antibody B-S12 from mouse ascites by anion exchange chromatography was established. The purity and recovery of B-S12 were up to 95% and 73% respectively. The purified antibody could stimulate the proliferation of XG-2 cells significantly in vitro.Conclusion:The method was characterized with high purity, excellent bioactivity and high recovery. It provided an important basis for its further application.
Ammonium sulfate precipitation
Ion chromatography
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AIM The purpose is to isolate and purify recombinant human interferon α2b. METHODS rHuIFNα2b expressed in engineering strain is isolated and purified by ultrasonic smash, ammonium sulfate precipitation, phosphoric acid acidification, cation exchange chromatography(CM-cellulose-32), anion exchange chromatograph (DEAE-cellulose-52) and gel ultrafiltration (Sephadex G-100). RESULTS The recovery of rHuIFNα2b was 30. The purified protein with a specific activity of 1.66×10+8 IU/mg appears as a peak with purity of 96 on the HPLC.=
Ammonium sulfate precipitation
Ultrafiltration (renal)
Sephadex
Ion chromatography
Phosphoric acid
Ammonium sulfate
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