| A method has been systematically developed to rapidly determine the major Cu, Fe and Zn-containing metalloproteins in rabbit plasma based on size exclusion chromatography and simultaneous on-line detection of Cu, Fe and Zn in the column effluent by an inductively coupled plasma atomic emission spectrometer. Whereas most previous studies reported on the analysis of serum, our investigations clearly demonstrated that the analysis of plasma within 30 min of collection results in the detection of one more Cu peak than has been previously reported. In contrast, only two Fe metalloproteins and ∼5 Zn metalloproteins were detected. Metalloproteins were assigned based on the co-elution of the corresponding metal and protein identified by immuno or activity-based enzyme assays. The developed SEC-ICP-AES approach allowed the determination of ∼12 Cu, Fe and Zn-metalloproteins in rabbit plasma and can be applied to analyze human plasma, which is potentially useful to diagnose Cu, Fe and Zn-related diseases. |
