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HPLC and tandem detection to monitor conformational properties of biopharmaceuticals
High-performance liquid chromatography (HPLC) with UV, circular dichroism (CD) and intrinsic fluorescence detection was applied to monitor conformational properties of recombinant human interferon alpha2b when performing size exclusion chromatography (SEC) and reversed-phase HPLC (RP-HPLC). In this...
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| Published in: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-07, Vol.821 (1), p.45-52 |
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| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c366t-74c9faccdaa25e3bd027a9a99ea808edb9adb2e8c51a519f7d242d1757b0f4e83 |
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| cites | cdi_FETCH-LOGICAL-c366t-74c9faccdaa25e3bd027a9a99ea808edb9adb2e8c51a519f7d242d1757b0f4e83 |
| container_end_page | 52 |
| container_issue | 1 |
| container_start_page | 45 |
| container_title | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences |
| container_volume | 821 |
| creator | LUYKX, Dion M. A. M GOERDAYAL, S. S DINGEMANSE, P. J JISKOOT, W JONGEN, Peter M. J. M |
| description | High-performance liquid chromatography (HPLC) with UV, circular dichroism (CD) and intrinsic fluorescence detection was applied to monitor conformational properties of recombinant human interferon alpha2b when performing size exclusion chromatography (SEC) and reversed-phase HPLC (RP-HPLC). In this way native conditions during SEC and structural changes of the protein during RP-HPLC were demonstrated. These results were confirmed by stand-alone fluorescence and CD measurements. With respect to HPLC tandem detection, the fluorescence detector compared favourably to the UV and CD detector regarding linearity, sensitivity and selectivity. SEC combined with intrinsic fluorescence scanning detection permits conformational analysis of small amounts of aggregates in the presence of excess native monomeric protein. In conclusion, HPLC with on-line UV and intrinsic fluorescence detection provides a promising concept for analysing the amount and conformational properties of a biopharmaceutical and its impurities. |
| doi_str_mv | 10.1016/j.jchromb.2005.04.005 |
| format | article |
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| ispartof | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2005-07, Vol.821 (1), p.45-52 |
| issn | 1570-0232 1873-376X |
| language | eng |
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| source | ScienceDirect Freedom Collection |
| subjects | Analysis Analytical, structural and metabolic biochemistry Biological and medical sciences Chromatography, Gel Chromatography, High Pressure Liquid - methods Circular Dichroism Fundamental and applied biological sciences. Psychology General pharmacology Humans Interferon-alpha - chemistry Interferon-alpha - isolation & purification Medical sciences Pharmaceutical Preparations - chemistry Pharmacology. Drug treatments Protein Conformation Recombinant Proteins Reproducibility of Results Sensitivity and Specificity Spectrometry, Fluorescence - methods Spectrophotometry, Ultraviolet |
| title | HPLC and tandem detection to monitor conformational properties of biopharmaceuticals |
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