Protein refolding in an oscillatory-flow-mixing reactor
We demonstrate that an oscillatory flow-mixing reactor (OFMR) operating in fed-batch mode is viable for industrial protein refolding. OFMR offers the advantage of uniform, controllable, and scalable mixing. Denatured lysozyme (15 mg/ml, in 8 M guanidine hydrochloride (GuHCl) or 8 M urea, and 32 mM...
| Main Authors: | , |
|---|---|
| Format: | Conference or Workshop Item |
| Published: |
2001
|
| Subjects: | |
| Online Access: | http://eprints.utm.my/3117/ |
| _version_ | 1848890509527875584 |
|---|---|
| author | Lee, C. T. Middelberg, Anton |
| author_facet | Lee, C. T. Middelberg, Anton |
| author_sort | Lee, C. T. |
| building | UTeM Institutional Repository |
| collection | Online Access |
| description | We demonstrate that an oscillatory flow-mixing reactor (OFMR) operating in fed-batch mode is viable for industrial protein refolding. OFMR offers the advantage of uniform, controllable, and scalable mixing. Denatured lysozyme (15 mg/ml, in 8 M guanidine hydrochloride (GuHCl) or 8 M urea, and 32 mM dithiothreitol) was fed into a 150-ml OFMR for 2 h and the final solution was incubated for a further 3 h. A control experiment used a small perfectly-mixed stirred-tank reactor (STR). Both systems were operated under mild and intense mixing conditions. With GuHCl denaturation, the refolding yield profiles throughout the 5 h period were identical in both the OFMR and STR, and yield was independent of mixing intensity. Conversely, refolding yield in both reactors following urea denaturation decreased from 25% to 15% as mixing intensity decreased. Clearly, imperfect mixing in large STRs may lead to reduced refolding yield when urea is used as a denaturant. This effect can be minimised by using a reactor with uniform, controllable and scaleable mixing, such as the OFMR. |
| first_indexed | 2025-11-15T20:43:12Z |
| format | Conference or Workshop Item |
| id | utm-3117 |
| institution | Universiti Teknologi Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T20:43:12Z |
| publishDate | 2001 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | utm-31172017-07-24T01:00:17Z http://eprints.utm.my/3117/ Protein refolding in an oscillatory-flow-mixing reactor Lee, C. T. Middelberg, Anton TP Chemical technology We demonstrate that an oscillatory flow-mixing reactor (OFMR) operating in fed-batch mode is viable for industrial protein refolding. OFMR offers the advantage of uniform, controllable, and scalable mixing. Denatured lysozyme (15 mg/ml, in 8 M guanidine hydrochloride (GuHCl) or 8 M urea, and 32 mM dithiothreitol) was fed into a 150-ml OFMR for 2 h and the final solution was incubated for a further 3 h. A control experiment used a small perfectly-mixed stirred-tank reactor (STR). Both systems were operated under mild and intense mixing conditions. With GuHCl denaturation, the refolding yield profiles throughout the 5 h period were identical in both the OFMR and STR, and yield was independent of mixing intensity. Conversely, refolding yield in both reactors following urea denaturation decreased from 25% to 15% as mixing intensity decreased. Clearly, imperfect mixing in large STRs may lead to reduced refolding yield when urea is used as a denaturant. This effect can be minimised by using a reactor with uniform, controllable and scaleable mixing, such as the OFMR. 2001 Conference or Workshop Item NonPeerReviewed Lee, C. T. and Middelberg, Anton (2001) Protein refolding in an oscillatory-flow-mixing reactor. In: The Proceeding of the 10th European Congress on Biotechnology, 8-11th July 2001, Madrid, Spain. |
| spellingShingle | TP Chemical technology Lee, C. T. Middelberg, Anton Protein refolding in an oscillatory-flow-mixing reactor |
| title | Protein refolding in an oscillatory-flow-mixing reactor |
| title_full | Protein refolding in an oscillatory-flow-mixing reactor |
| title_fullStr | Protein refolding in an oscillatory-flow-mixing reactor |
| title_full_unstemmed | Protein refolding in an oscillatory-flow-mixing reactor |
| title_short | Protein refolding in an oscillatory-flow-mixing reactor |
| title_sort | protein refolding in an oscillatory-flow-mixing reactor |
| topic | TP Chemical technology |
| url | http://eprints.utm.my/3117/ |