i am planning to extract silk fibroin proteins from cocoon. can anyone please suggest me the easiest and less expensive way for extracting silk fibroin protein?
Hi Pragyan,
The selection of the right method I think depend on the desired properties of the fibroin.
If for example the plan is to use fibroin for biomedical application then the purity of fibroin is critical and residual of sericin proteins can trigger an immunogenic response in vivo. The hydrolysis of sericins can be performed at room temp to 100 °C using alkali acids (e.g. sodium carbonate and citric acid) or using denaturing agent such as urea at high concentration. Nevertheless, the alkali treatment is a common technique.
In this method, cocoons are boiled in a sodium carbonate solution (the molarity and time can effect on the degree of hydrolysis of fibroin and therefore its MW). Then the fibroin can be dissolved in salt solution such as Libr or a calcium chloride/water/ethanol system. Lastly, dialysis can be used to remove the salts to obtain a fibroin solution.
I hope it will help
Hi Pragyan,
The selection of the right method I think depend on the desired properties of the fibroin.
If for example the plan is to use fibroin for biomedical application then the purity of fibroin is critical and residual of sericin proteins can trigger an immunogenic response in vivo. The hydrolysis of sericins can be performed at room temp to 100 °C using alkali acids (e.g. sodium carbonate and citric acid) or using denaturing agent such as urea at high concentration. Nevertheless, the alkali treatment is a common technique.
In this method, cocoons are boiled in a sodium carbonate solution (the molarity and time can effect on the degree of hydrolysis of fibroin and therefore its MW). Then the fibroin can be dissolved in salt solution such as Libr or a calcium chloride/water/ethanol system. Lastly, dialysis can be used to remove the salts to obtain a fibroin solution.
I hope it will help
Amin Shavandi: Thank you sir.. this was very helpful. i am planning to use silk fibroin protein for biomaterials applications. i am planning to dope with hydrogel.
Hi
For isolation of silk fibroin from cocoons you can refer these papers,
Impact of silk fibroin-based scaffold structures on human osteoblast MG63 cell attachment and proliferation
Silk–PVA Hybrid Nanofibrous Scaffolds for Enhanced Primary Human Meniscal Cell Proliferation
All the best
Mamatha M Pillai : Thank you mam for the reference. It going to be helpful for my work.
Hi Amin Shavandi,
Thank you very much for your useful answer. Could you please tell the detail ratio of calcium chloride/water/ethanol system for dissolving the fibroin. Do we need the dialysis cassette for the dialysis to remove the salts to obtain a fibroin solution? Is there any method without using dialysis cassette?
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