Analytical size exclusion chromatography of Hbs with or without Hp.

<p>A. Representative chromatograms of Hb, ααXLHb, and ββXLHb in the presence or absence of Hp. The absorbance was monitored at 405 nm for heme, and the peaks appearing between the elution time 13 and 19 minutes are Hb and Hp complexes. B. The percentage of Hp bound with Hb was determined and compared among ααXLHb, ββXLHb, and Hb based on baseline normalized area under curve for peaks within the boxed region of the eluting profiles (from Panel A). Statistical analysis was performed using a One-way analysis of variance with a Bonferroni’s Multiple comparison test to evaluate between group differences. Data are represented as mean areas +/− sem, significance was set at p<0.05. All analysis were performed using GraphPad Prism software.