Immunoaffinity beads for selective removal of cholesterol from human plasma

Anti-low density lipoprotein antibody (anti-LDL antibody) attached poly(2-hydroxyethyl methacrylate-methacryloylamidophenylalanine) (poly(HEMA-MAPA)) beads were prepared for selective removal of cholesterol from hypercholesterolemic human plasma. Poly(HEMA-MAPA) beads were produced by a modified suspension polymerization and then characterized by swelling tests and SEM. Blood-compatibility tests were also investigated: The water swelling ratio of the poly(HEMA-MAPA) beads increased significantly (68%) compared with pHEMA (55%). All the clotting times increased when compared with poly(HEMA) beads. Loss of platelets and leukocytes was very low. The maximum anti-LDL antibody attachment was achieved at pH 7.0. Attachment of anti-LDL antibody was 29.6 mg/g. There was a very low non-specific cholesterol binding onto the poly(HEMA-MAPA) beads, about 0.74 mg/g. Anti-LDL antibody attached beads adsorbed in the range of 13.3-16.0 mg cholesterol/g from hypercholesterolemic human plasma. Up to 92% of the adsorbed LDL was desorbed. The binding-elution cycle was repeated 10 times using the same beads. There was no significant loss of binding capacity.