New Ligand Coupling Procedure for Formation of an Immunoadsorption Wall

To prevent the occurrence of dialysis-related amyloidosis, an immunoadsorption wall based on polyacrylamide has been manufactured by a recently developed, partially incomplete, two-stage polymerization method. During the preparation process, efficient utilization of coupling antibodies is the key to large-scale production of such a toxin removal modality. In this study, we attempted to carry out the ligand coupling procedure after formation of a cyanogen bromide (CNBr)-activated stationary phase, using anti-beta2-microglobulin (&bgr;-2M) antibodies. In vitro immunoadsorption tests show that the levels of &bgr;-2M decrease rapidly within the first 2 hours for all the immunoadsorption tests. After that, nearly blank values were reached for tests of initial levels of c. 30 &mgr;g/mL and c. 82 &mgr;g/mL, whereas a relatively constant level of c. 10 &mgr;g/mL was maintained for the test of initial levels of c. 185&mgr;g/mL. The maximum surface binding capacity of the prepared immunoadsorption walls is estimated by fitting experimental data, using a mathematical model of saturation kinetics. The present comparative investigation also suggests the manufacturing process for an immunoadsorption wall could be improved and facilitated by this new ligand coupling procedure without compromising the resulting binding capacity. Furthermore, the experimental protocols as well as the present methodology could be helpful for development of a clinically applicable immunoadsorption wall.