title: Ex vivo biocompatibility of avidin-agarose: a new device for direct adsorption of biotinylated antibodies from human whole blood.
abstract: Radioimmunotherapy using radiolabeled antitumor antibodies (RAA) is limited by the toxicity of unbound antibodies in the circulation. Removal of excessive antibodies by affinity-adsorption could therefore allow the administration of increased dosages of RAA while decreasing their adverse effects. Recently, avidin-agarose (AA) minicolumns were used in animal experiments for the removal of biotinylated antibodies from whole blood exploiting the high affinity binding of biotin to avidin (pK 1015 M-1). This study was performed to evaluate the ex vivo biocompatibility of AA minicolumns with human blood. Ten ml AA minicolumns were perfused online ex vivo in the single pass mode with fresh blood from 8 healthy donors at a flow rate of 6.25 ml/min. The anticoagulation consisted of 0.5 IU heparin plus 0.0-2.1 mg citrate per ml of blood. In Part 1 of the study (40 min perfusion, n = 4), the optimal anticoagulation was found to be 0.5 IU heparin plus about 1 mg citrate per ml of blood. In Part 2 of the study, four 80 min test-runs were performed. No signs of hemolysis were found, and the thrombogenicity of the AA gel was negligible. Cell counts and column inlet pressures remained constant; toward the end of the 80 min test-runs, some activation of blood cells (elastase, beta-thromboglobulin), the complement system (C3a, C5a) and the plasmatic coagulation (thrombin-antithrombin complex) was detectable. A moderate initial bradykinin release rapidly subsided to very low levels. In summary, AA minicolumns showed good biocompatibility upon contact with human whole blood and merit further investigation in a closed-loop system for a potential application of direct tumor antibody removal by hemoperfusion.
authors.affiliation: Nephrology Division, Medical Clinic I, Klinikum Grosshadern, University of Munich, Munich, Germany. bosch@med1.med.uni-muenchen.de
authors.forename: T
authors.initials: T
authors.lastname: Bosch
authors.forename: A
authors.initials: A
authors.lastname: Lennertz
authors.forename: C
authors.initials: C
authors.lastname: Duhr
authors.forename: E
authors.initials: E
authors.lastname: Fink
authors.forename: W
authors.initials: W
authors.lastname: Samtleben
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000097:blood*; D000912:Antibodies, Neoplasm / Q000097:blood; D000925:Anticoagulants / Q000097:blood; D000990:Antithrombin III; D001360:Avidin / Q000737:chemistry*; D001672:Biocompatible Materials / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D001772:Blood Cell Count; D001920:Bradykinin / Q000097:blood; D002846:Chromatography, Affinity; D002951:Citrates / Q000097:blood; D015926:Complement C3a / Q000032:analysis; D015936:Complement C5a / Q000032:analysis; D006461:Hemolysis; D006464:Hemoperfusion / Q000295:instrumentation / Q000379:methods*; D006493:Heparin / Q000097:blood; D006801:Humans; D008024:Ligands; D010196:Pancreatic Elastase / Q000097:blood; D010447:Peptide Hydrolases; D011312:Pressure; D016499:Radioimmunotherapy; D012685:Sepharose / Q000737:chemistry*; D013927:Thrombosis / Q000517:prevention & control; D001620:beta-Thromboglobulin / Q000032:analysis
chemical_list: D000906:Antibodies; D000912:Antibodies, Neoplasm; D000925:Anticoagulants; D001672:Biocompatible Materials; D002951:Citrates; D008024:Ligands; C046193:antithrombin III-protease complex; D001620:beta-Thromboglobulin; D001360:Avidin; D001710:Biotin; D015926:Complement C3a; D015936:Complement C5a; D000990:Antithrombin III; D006493:Heparin; D012685:Sepharose; D010447:Peptide Hydrolases; D010196:Pancreatic Elastase; D001920:Bradykinin