Continuous beds. Their applicability for immobilization of proteins

Abstract

An epoxy-activated continuous bed can be prepared for immobilization of proteins in a simple, rapid, and cost-effective way. The concentration of epoxy groups on the continuous bed was as high as 600 mumol/mL compressed bed (compression of the bed decreases the peak broadening). Human transferrin, human serum albumin and particularly urease were employed as model proteins. The immobilization of urease was virtually completed within 1 h in 1 M potassium phosphate, pH 7.4. The binding capacity was 97 mg of urease/mL compressed bed. This bed is of clinical interest, since it is inexpensive to prepare and permits reproducible enzymatic determination of urea in serum and urine (the chromatographic step is finished within 1-2 min).