Non-denaturing assay for the determination of the potency of recombinant bovine somatotropin by high-performance size-exclusion chromatography
| Autor: | Ghrist Bf, Tucker Rc, Coleman Mr, Jen P. Chang |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Protein Denaturation
Base (chemistry) Size-exclusion chromatography Biochemistry Analytical Chemistry law.invention chemistry.chemical_compound law Potency Animals Bovine somatotropin Ammonium Agrégation Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography Chemistry Organic Chemistry General Medicine Polymer Recombinant Proteins Rats Molecular Weight Solubility Growth Hormone Recombinant DNA Chromatography Gel Cattle Spectrophotometry Ultraviolet |
| Zdroj: | Journal of chromatography. A. 675(1-2) |
| ISSN: | 0021-9673 |
| Popis: | A non-denaturing high-performance size-exclusion chromatographic method has been developed for the determination of the potency of recombinant bovine somatotropin (rbST) in bulk materials. A Spherogel TSK 3000 PW column containing a polymer base packing material with very hydrophilic bonded surface was used in this method. Ammonium hydrogencarbonate buffer pH 9.0, was used as the mobile phase. This method was shown to be non-denaturing by rat mass-gain assay and radioreceptor assay. The optimization for the separation and determination of rbST has been investigated. The method was validated for the determination of rbST in bulk materials. In addition, rbST soluble aggregates formed in the production process due to protein association, used to be found in bulk materials. The behavior of rbST soluble aggregates in ammonium hydrogencarbonate solutions have been studied. The bio-inactive aggregates can be separated by the method developed in this study. The high—low chromatographic technique has been used to estimate rbST soluble aggregates in bulk materials. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|