| Full text | |
| Author(s): |
Dias, Paulo V. S.
[1]
;
Arthuso, Fernanda S.
[1]
;
Oliveira, Joao E.
[1]
;
Suzuki, Miriam F.
[1]
;
Sousa, Jose M.
[1]
;
Ribela, Maria Teresa C. P.
[1]
;
Bartolini, Paolo
[1]
;
Soares, Carlos R. J.
[1]
Total Authors: 8
|
| Affiliation: | [1] CNEN SP, IPEN, Biotechnol Ctr, Sao Paulo - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES; v. 1072, p. 193-198, JAN 1 2018. |
| Web of Science Citations: | 1 |
| Abstract | |
Reversed-phase high-performance liquid chromatography (RP-HPLC) has been used to analyze Interferon alpha-2 (IFN-alpha 2) as a pure protein or as a pharmaceutical preparation: a method for analyzing periplasmic IFN-a2 directly in osmotic shock extract has, however, never been reported. This work describes an RP-HPLC methodology for the qualitative and quantitative analysis of human IFN-alpha 2a and IFN-alpha 2b directly in bacterial periplasmic extracts or in purified preparations. The analytical method has been set up and validated for accuracy, precision, linearity, sensitivity and specificity. A recovery test indicated an average bias of similar to 1%, intra-day and inter-day quantitative determinations presented relative standard deviations always <= 5%, while the working sensitivity was of similar to 0.3 mu g of IFN-alpha 2 (RSD = 5%). The method proved to be suitable for detecting and quantifying also glycosylated and oxidized forms and N-methionylated IFN-alpha 2 molecules, it was, however, not able to distinguish between IFN-alpha 2a and IFN-alpha 2b. This rapid methodology allows the application of RP-HPLC as a powerful tool to monitor the production yield and quality of IFN-alpha 2 in osmotic shock fluids, right after, or even during the fermentation process. (AU) | |
| FAPESP's process: | 15/10928-6 - Development of fermentation in a bioreactor for the production of interferon ±-2a secreted into the periplasmic space of Escherichia coli |
| Grantee: | Paulo Victor Sarmento Dias |
| Support type: | Scholarships in Brazil - Master |
| FAPESP's process: | 15/26058-0 - Glycoprofiling and N-glycosylation site occupancy of different human thyrotropin (hTSH) preparations of pituitary or recombinant origin |
| Grantee: | Carlos Roberto Jorge Soares |
| Support type: | Regular Research Grants |
| FAPESP's process: | 12/24345-4 - Evaluation of the potential of prolactin antagonists to the treatment of obesity and Diabetes mellitus 2 |
| Grantee: | Carlos Roberto Jorge Soares |
| Support type: | Regular Research Grants |