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. Author manuscript; available in PMC: 2022 May 29.
Published in final edited form as: Bioconjug Chem. 2019 Feb 28;30(3):568–571. doi: 10.1021/acs.bioconjchem.9b00130

Figure 3.

Figure 3.

(A) RP-HPLC analysis of SS lysate was performed after incubation with NaBH3CN. (B) The relationship of GBT levels and the retention fraction of subunit globins. The growing fractions of GBT-Hb adducts were also plotted in the figure (n = 3). The fractions of all the species were normalized by the globin chain area compared to the heme area (calculated from the Figure 3A). (C−F) Intact mass spectrometric analysis of Hb subunits from patient hemolysate (including control) treated with increasing dosages of GBT440. (G) LC/MS/MS fragmentation spectrum representing y and b fragment ions for the GBT440 labeled N-terminal α peptide. The GBT440 concentrations 30, 70, 120, and 150 correspond to % values of the 1 mM HbS tetramer (i.e., SS 30 contains 0.3 mM GBT440). These mass spectrometry data confirm that GBT440 labeling is specific to the α N-terminus.