Fig. 7
Size distribution of low molecular weight AGE-peptides from diabetic patients on hemodialysis.
Low molecular weight fractions obtained from diabetic patients with end-stage renal disease on hemodialysis were subjected to gel filtration chromatography (Sephadex G-15 column: 1.5 × 110 cm). The eluted fractions were monitored for non-CML AGE (glyceraldehyde, ◯; glycolaldehyde, ▲; methylglyoxal, ■; glyoxal, ◊) by ELISA (data from triplicate determinations), AGE florescence (Ex = 360 nm/Em = 440 nm, A) and absorbance at 280 nm (●). Elution of cytochrome c (V0), vitamin B12 (1,355 Da) and cytidine (243 Da) (molecular weight markers) is indicated by arrows. (A) Chromatography monitored by AGE fluorescence and absorbance at 280 nm. (B) Chromatography with affinity-purified non-CML AGE antibodies obtained by incubation of AGE-RSA with glyceraldehyde and glycolaldehyde. (C) Chromatography with affinity-purified non-CML AGE antibodies obtained by incubation of AGE-RSA with methylglyoxal and glyoxal.