System and methods for scalable melanin purification
Provided are systems and methods for isolating melanins from a medium that include providing tangential filtration of the medium across a membrane with a 300 kilo Dalton or less cutoff. In some aspects, the methods and systems are suitable for large scale preparations due to the inherent established scalability of the unit operations employed herein. The systems and the methods can be utilized to purify melanins from fermented cell medium as well as from lysates generated therefrom.
1 . A method to isolate one or more melanins from an aqueous medium, the method comprising:
providing the aqueous medium comprising the one or more melanins therein to an inlet of a tangential flow filtration (TFF) enclosure at a positive pressure, wherein the TFF enclosure comprises a retentate outlet and a permeate outlet; and
flowing the aqueous medium across a first membrane within the TFF enclosure toward the retentate outlet, wherein the membrane has a size restriction of about 300 kilo Daltons (kDa) or less to isolate the one or more melanins from the aqueous medium, and wherein the permeate outlet is operably connected to a permeate storage container for collecting permeate.
2 . The method of claim 1 , wherein a pump is placed in front of the inlet to provide the positive pressure.
3 . The method of claim 1 , wherein the positive pressure is of about 1 to about 10 pounds per square inch (PSI).
4 . The method of claim 1 , wherein the retentate outlet is operably connected to the inlet to allow the aqueous medium to recirculate through the TFF enclosure.
5 . The method of claim 1 , wherein the inlet is operably connected to a retentate storage container.
6 . The method of claim 5 , wherein the retentate outlet is operably connected to the retentate storage container.
7 . The method of claim 1 , further comprising one or more additional membranes operably connected to the first membrane.
8 . The method of claim 1 , wherein the aqueous medium comprises a fermentation broth.
9 . The method of claim 1 , wherein the aqueous medium comprises cellular lysate.
10 . The method of claim 9 , wherein the cellular lysate is pretreated with at least one of a DNAse, a RNAse, or a proteinase.
11 . The method of claim 1 , further comprising drying a permeate collected within the permeate storage container.
12 . The method of claim 1 , further comprising cleaning the TFF enclosure prior to providing the aqueous medium.