Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 1209, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2022.123410
Keywords
Inclusion bodies; HCK; Chaperoning system; On -column refolding
Funding
- National Natural Science Foundation of China [30800188]
- Special Funds for Co-construction Project of Beijing Municipal Commission of Education
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This article describes a novel method for the renaturation and purification of inclusion bodies, which significantly increases the activity recovery of bioactive proteins.
The formation of inclusion bodies in bacterial hosts poses a major challenge for the large-scale recovery of bioactive proteins. The process of obtaining bioactive protein from inclusion bodies is labor intensive, and the yields of recombinant protein are often low. Here, we describe a novel method for the renaturation and puri-fication of inclusion bodies. This method combines a scFv-oligopeptide chaperoning system and an on-column refolding system to help refold human muscle creatine kinase (HCK) inclusion bodies. This method could significantly increase the activity recovery of denatured HCK inclusion bodies and provides an effective method for the production of bioactive proteins from inclusion bodies.
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