Fig. 7
Size distribution of AGE in serum from diabetic patients on hemodialysis. Serum from diabetic patients with end-stage renal disease on hemodialysis was subjected to gel filtration chromatography (Sephacryl S-200 column, 1.5 × 110 cm). Eluted fractions were monitored for non-CML AGE (○) and CML (●) by ELISA (data from triplicate determinations), AGE fluorescence (△: Ex = 360 nm/Em = 440 nm), and absorbance at 280 nm (▲). Elution of aldolase (160,000 Da), BSA (67,000 Da), chymotrypsinogen A (25,000 Da), and vitamin B12 (1355 Da) (molecular weight markers) is indicated by arrows. (A) Chromatography monitored by AGE fluorescence and absorbance at 280 nm. (B) Chromatography with affinity-purified CML or non-CML AGE antibodies obtained using 12-week or 8-week AGE-RSA, respectively.