I'am not a hundred percent shure on this, since I dont have practical experience with this enzyme, but i would suggest there are two main possibilities:
1. The crude collagenase gontains a small amount of other similar digestive enzymes and is thus more effective for digesting cells but posssibly less effective in digesting ONLY the collagen.
2. The enzyme is partially damaged during the purification process, decreasing the overall activity per molecule.
Crude collagenase has other proteases and perhaps other hydrolytic enzymes that will also act to disaggregate cells and ECM materials.
possibly also crude enzyme has a lot of protein impurities. If these bind non specifically to the plastic of the flask then more collagenase remains in solution to be active but in the case of pure collagenase binding to the plastic will remove good enzyme from the digesting solution
Thank you for the answers :)
These do make sense. However, would it not be unadvisable to use crude collagenase for the process ?
Is it a possibility that the contaminants would somehow affect the cell growth negatively ?
A possible advantage of using the crude collagenase is that your dissociation procedure will take less total time, and thus the cells may be less stressed, as they are returned sooner to the comfort of their culture medium. Of course, this assumes that after the disaggregation incubation, you do an adequate job of rinsing the collagenase solution away from the cells.
- Badges
- Science method