Cross-linked cellulose membranes
View Patent ↗The invention relates to a process for making porous cross-linked charged cellulosic polymeric membranes capable of binding to a target molecule. The invention provides methods for separating target molecules from other components in a solution comprising use of membranes obtainable by the process of the invention. The method has particular utility in separating proteins and nucleic acids from cell lysates and cultures.
1. A process for making a porous cross-linked charged cellulosic polymeric membrane comprising:
i) adding a base to a porous cellulose acetate membrane comprising a plurality of acetate groups in the presence of a solution of 0.25-2% epichlorohydrin to cross-link said cellulose acetate therein to provide a cross-linked cellulose acetate membrane, and
ii) reacting said cross-linked membrane with glycidyl trimethylammonium chloride to introduce a positively charged group
wherein said process is carried out in the presence of sodium sulfate, and the resulting porous cross-linked charged cellulosic polymeric membrane comprises a plurality of cellulose acetate groups and swells from 40% to 250% by volume in a solution of water.
2. The process of claim 1 , wherein the resulting cross-linked cellulosic membrane swells by 125% to 250% by volume in a solution of water.
3. The process of claim 1 , wherein said base is selected from the group consisting of sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, tetraalkylammonium hydroxide, sodium carbonate, calcium carbonate, sodium triphosphate, sodium silicate, potassium carbonate, potassium silicate, potassium triphosphate and sodium hydrogen carbonate.
4. The process of claim 1 , wherein the membrane comprises pores having a pore size of 0.1 to 10 microns.
5. The process of claim 1 , wherein the base is sodium hydroxide.
6. The process of claim 1 , wherein the crosslinking process is carried out at 20° C. to 60° C. for a period of at least 1 hour.
7. The process of claim 6 , wherein the crosslinking process is carried out at a temperature of 45° C. to 55° C. for a period of at least 1 hour.
8. The process of claim 1 , wherein said ligand comprises a positively charged group and wherein the resulting porous cross-linked charged cellulosic polymeric membrane has a dynamic BSA capacity of at least 30 mg/ml.