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Volume 6, Issue 4 (Suppl)
Clin Pharmacol Biopharm, an open access journal
ISSN: 2167-065X
Page 93
Euro Biopharma & Ethnopharmacology 2017
November 09-11, 2017
&
6
th
International Conference and Exhibition on
November 09-11, 2017 Vienna, Austria
4
th
EUROPEAN BIOPHARMA CONGRESS
PHARMACOLOGY AND ETHNOPHARMACOLOGY
Joint Event
Refolding control of highly disulfide bonded proteins by multi gradient on-column strategy
Syed Babak Mousavi
and
Ahmad Fazeli
Zistdaru Danesh Co. Ltd., Iran
C
orrect formation of disulfide bonds is crucial in the refolding of multi disulfide bonded inclusion body proteins. Covalent
aggregation due to wrong cross linking of disulfide bonds leads to a low refolding yield. On-column refolding can be used
to refold complex proteins under controlled conditions. This study reports the development of a multi gradient strategy for
on-column refolding of high disulfide bonded proteins using size exclusion chromatography. Recombinant tissue plasminogen
activator (r-PA, reteplase) with 9 disulfide bonds was used as the protein model. Applying a four-gradient strategy, including
decreasing linear gradient of urea concentration and increasing linear gradients of pH, cysteine, and arginine concentration at
the same time, resulted in 49.72% activity recovery and 91% mass recovery, which was 4 and 2.5-fold more than conventional
on-column refolding with non-gradient and urea-arginine gradient strategies respectively. Successful application of multi
gradient strategy in improving refolding yield of reteplase demonstrated controllability of the refolding process using size
exclusion chromatography with a rationally designed gradient strategy which can be used in the refolding of other complex
proteins too.
babak.musavy@gmail.comClin Pharmacol Biopharm 2017, 6:4(Suppl)
DOI: 10.4172/2167-065X-C1-026