Specificity of Anti-Amadori hCD59 Antibodies

Example 3

The specificity of anti-Amadori-modified hGCD59 mAb D2 and D3 to glycated hCD59 was tested in cell lysates derived from diabetic transgenic mice expressing hCD59 transgene (Tg) in red blood corpuscles (RBCs). Diabetes was induced in the hCD59 transgenic mice by administering one dose of Streptozotein (STZ) and blood glucose was measured after two weeks. A mouse was considered diabetic if its blood sugar level was greater than 200 mg/dL. RBCs were obtained from diabetic transgenic mice (D) and control non-diabetic mice (ND), lysed and proteins were extracted. The protein samples were separated using SDS-PAGE (Sodium Dodecyl sulfate-Polyacrylamide gel electrophoresis) and immunoblotted with anti-Amadori modified hGCD59 antibody (D2) and anti hCD59 antibody (FIG. 2). Consistent with the elevation of glycated CD59 in diabetes, the anti-Amadori modified hGCD59 antibody showed an intense band in the diabetic mice and only a faint band in the control non-diabetic mice. Both the diabetic and control non-diabetic mice showed similar levels of hCD59, demonstrating the specificity of the anti-Amadori modified hGCD59 antibody.