In the current investigation, the expression yield and solubility of aβ antibody was analyzed using t7 promoter in fed-batch cultivation mode. pcr amplification of aβ was carried out using the specific primersand cloned in the hi-control 10g cells of lucigen kit. after transforming the cloned gene, the recombinant escherichia coli10g cells were cultivated in batch and fed-batch culture medium. the expression and solubility of aβ was investigated in two different temperatures (25 and 30°c) and different induction time(4,6,8,12,24 hours). protein expression was analyzed by sds-page and western blotting technique.
Methods
Aggregation of amiloiyd beta (aβ) is the most important reasons of alzheimer’s disease(ad). passive immunotherapy with monoclonal antibodies against aβ is one of the subjects of recent studies to prevent ad. recombinant protein expression technology is the most important approach for production of biomolecules. escheriachia coli is one of the most popular hosts for protein expression in biotechnology laboratories. inspite its simplicity, expressed proteins are mostly in insoluble form, called inclusion bodies. several strategies can overcome this problem. changing the growth medium and co-expression of chaperones are some of these approaches.
Results
the maximum expression of protein and cell density was observed in enbase cultivation system.the solubility of expressed protein was slightly increased in enbase medium specially at the initial 6hours after induction.
Conclusion
Therefore, due to slow and continuous release of glucose and controlled growth, a highly protein expression of aβ was obtained in fed-batch cultivation mode. longer time of expression give protein the enough time for folding correctly and increase solubility.
