The journal article entitled "Potential of sequential treatment with minocycline and S. aureus specific phage lysin in eradication of MRSA biofilms: an in vitro study" by Chopra et al. (2015) mentioned following a method described by Adams and Park (1956).
Many endolysins are thermostable (not all of them) and this is something that you can use. Take the whole extract after the phage inducing (I would do it in a buffer and not in the culture medium). Centrifuge the extract around 12000- 15 000 g for 10 min at +4. Recover the supernatant and heat it at 95 oC during 10 minutes in a thermoblock or a water bath. Transfer the tubes to ice for one minute and immediately centrifuge the extract around 12000- 15 000 g for 10 min at +4. The supernatant should be endolysins rich and should have antibacterial activity (sometimes is only species/strain specific). You can increase the activity by concentrating the extract in a centicon devise or similar (http://kirschner.med.harvard.edu/files/protocols/Millipore_Centricons.pdf). However, if yuo need something pure you should proceed to some chromatographic steps, maybe with a Gel filtration after concentration should be OK. I hope this helps.
u may discuss the issue with Dr. Santi M. Mandal, Assistant Professor , Vidyasagar University, email: [email protected]
Dear Katherine, you can follow the links below which may help you to obtain certain crucial informations regarding your topic:-
1. http://www.ncbi.nlm.nih.gov/pubmed/22475951
2. http://www.bio-protocol.org/e1208
Hello and good time, Mrs Katherine.
I recommend that use from size-exclusion chromatography (SEC) to purification Endolysin (of course, first you should to induce your phages)
The best method is to clone and express your gene of interest. It will reduce the cost as well as efforts for subsequent purifications in long runs. Endolysins to be used for in-vivo applications must be endotoxin-free, so you have to keep enough protein with you to bear the loss during endotoxin removal, which can be achieved by overexpression only. Conventional methods like ion exchange chromatography may be cheaper than overexpression at a point but the yield is very low to be used for repetitive experiments. So I recommend you to just clone it once and express for years!!!!!
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