Patent ID: 9291619
Filing Date: 2016-03-22
CPC Classification: G01N

Claim Text:
1. A method of quantifying a ligand-bound fraction of redox-active non-transferrin bound iron (NTBI) in a biological fluid, the method comprising: (a) quantifying enhanced labile plasma iron (eLPI) in a first sample of the biological fluid, wherein said eLPI consists of both ligand-bound and non-ligand-bound redox-active NTBI iron, comprising: (i) contacting a sample of the biological fluid with a reducing agent to obtain redox-active iron; (ii) contacting said sample of the biological fluid with a nitrilotriacetate (NTA) at a concentration of 0.4-2 mM to mobilize ligand-bound redox-active non-transferrin bound iron in said sample; (iii) contacting said sample of the biological fluid of step (a)(ii) with an indicator including an iron binding moiety and fluorophore; and (iv) detecting and quantifying a fluorescent signal of said fluorophore thereby quantifying the eLPI in the biological sample, (b) quantifying the labile plasma iron (LPI) in a second sample of the biological fluid, wherein said LPI consists of non-ligand bound redox-active non-transferrin bound iron NTBI, comprising: (i) contacting said second sample of the biological fluid with a reducing agent to obtain redox active iron; (ii) contacting said second sample of the biological fluid of step (b)(i) with an indicator including an iron binding moiety and fluorophore; and (iii) detecting and quantifying a fluorescent signal of said fluorophore thereby quantifying the LPI in the biological sample, and (c) subtracting the LPI from eLPI, thereby quantifying the ligand-bound fraction of redox-active non-transferrin bound iron (NTBI) in the biological fluid.