Towards mechanized biparametric ceruloplasmin assay

Area of Science:

• Analytical Chemistry
• Biochemistry
• Clinical Diagnostics

Background:

• Ceruloplasmin (Cp) is a crucial metalloprotein involved in iron transport and antioxidant defense.
• Accurate determination of Cp activity and concentration is vital for diagnosing various diseases.
• Existing analytical methods may lack efficiency or require complex procedures.

Purpose of the Study:

• To develop and validate two automated Multicommutated Flow Analysis (MCFA) systems.
• To enable simultaneous determination of both biocatalytic activity and concentration of ceruloplasmin.
• To optimize analytical parameters for enhanced sensitivity and efficiency in human serum analysis.

Main Methods:

• Design of two distinct MCFA systems utilizing solenoid pumps and valves.
• Kinetic measurements using p-phenylenediamine and hydrogen peroxide for catalytic activity assay.
• Light-scattering detection coupled with immunoprecipitation for concentration determination.

Main Results:

• The developed flow bioanalytical system for Cp activity achieved LOD of 0.07 U/mL and LOQ of 0.38 U/mL with RSD ≤6%.
• The MCFA system for Cp concentration provided 35 measurements/hour with LOD of 0.9 mg/L and LOQ of 3.2 mg/L.
• Both systems demonstrated successful application in analyzing real human serum samples.

Conclusions:

• The proposed automated MCFA systems provide versatile and efficient platforms for ceruloplasmin analysis.
• These methods offer improved analytical performance, including sensitivity and speed, for clinical diagnostics.
• The developed systems are suitable for routine analysis of human serum samples.