Methacrylated Heparin Sulfate
1. In a 250 mL round bottom flask, dissolve 5g (7.634 mmol unit, unit MW = 665) Heparin sodium salt in a 200 mL buffer solution of 50 mM (1.952g) 2-morpholinoethanesulfonic acid (MES) and 0.5 M (5.8443g) NaCl.
2. Prepare appropriately diluted NHS, EDC and AMEA solutions. (Dilute in MES)
3. Add to the solution 0.439 g (3.817 mmol) N-hydroxysuccinimide (NHS) and 1.463 g (7.634 mmol) 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) (molar ratio of HS (units):NHS:EDC = 1:0.5:1) , stir the solution for 5 minutes.
4. Add 0.632 g 2-aminoethyl methacrylate (AEMA) (molar ratio of NHS:EDC:AEMA = 1:2:1) to the solution and maintain the reaction at room temperature for 24 h.
5. Purify the product solution using dialysis tubing (12-14kDa MWCO) against a 4L, 25 C DI water bath. Keep the water bath stirred by stir bar. Replace DI water every 24 hours for 4 days.
6. Lyophilize the purified product. Store the product at -20 C.