Tracking Heme-Protein Interactions in Healthy and Pathological Human Serum in Native Conditions by Miniaturized FFF-Multidetection

The interaction of heme with blood serum proteins plays an important role in many physiological and pathological processes involving enzyme activity, gene expression cell proliferation. mechanisms underlying these interactions are; however, not yet fully understood. New analytical methods able to investigate protein-heme binding native, biologically representative conditions are thus required. In this work, we present a method based on miniaturized, hollow-fiber flow field-flow fractionation multiple spectrophotometric light-scattering detection for size separation high-abundance selective heme-bound subpopulations. Heme is found mainly interact albumin, whereas low amount also binds other such as IgM. ability bind investigated individual proteins. IgG unable at clinically relevant concentrations. proposed allows separation, quantitation, mass/size characterization proteins, providing information heme-protein complex stability preferred heme-clearing pathways. same approach could be perspective extended the investigation specific heme-antibody binding, further studies molecules pharmaceutical/clinical interest.