Assessment of the stability of an immunoadsorbent for the extracorporeal removal of Beta-2-microglobulin from blood.
BACKGROUND
Dialysis-related amyloidosis (DRA) is a devastating and costly condition that affects patients with end stage kidney disease. A key feature of DRA is the formation of amyloid fibrils, consisting primarily of beta2-microglobulin. Except for kidney transplantation, conventional kidney replacement therapies, which are based on nonspecific mechanisms, do not adequately address beta2-microglobulin removal. An antihuman beta2-microglobulin single-chain variable region antibody fragment (scFv) was developed to confer specificity to beta2-microglobulin removal during hemodialysis.


METHODS
The scFv was immobilized onto agarose and characterized for beta2m binding capacity, thermal stability at 37 degrees C, regeneration capacity, storage conditions, and sterility.


RESULTS
The beta2-microglobulin binding capacity was 1.3 mg/ml scFv gel. The immunoadsorbent is thermally stable, can be regenerated, stored short-term in 20% ethanol, lyophilized for long-term storage, and withstand process conditions similar to that of a patient's hemodialysis therapy.


CONCLUSIONS
The results support further investigation of immobilized scFvs as a novel tool to remove beta2-microglobulin from blood.
Daniels
Cynthia M
CM
Biomedical Engineering Department, Northwestern University, Evanston, IL 60208, USA.
Woolverton
Emily M
EM
Sprague
Stuart M
SM
Ameer
Guillermo A
GA
D000686:Amyloidosis / Q000209:etiology / Q000517:prevention & control*; D004355:Drug Stability; D005112:Extracorporeal Circulation; D006801:Humans; D007135:Immunoglobulin Variable Region; D007163:Immunosorbent Techniques; D007676:Kidney Failure, Chronic / Q000150:complications / Q000628:therapy*; D006435:Renal Dialysis / Q000009:adverse effects; D012685:Sepharose; D013242:Sterilization / Q000379:methods; D001613:beta 2-Microglobulin / Q000276:immunology* / Q000302:isolation & purification*
D007135:Immunoglobulin Variable Region; D001613:beta 2-Microglobulin; D012685:Sepharose