Objective To establish and evaluate a protein microarray method for combined measurement of serum ferritin (SF) and soluble transferrin receptor (sTfR).Methods Microarrayer was used to print both anti-SF antibodies Ⅰ and anti-sTfR antibodies I on each protein microarray.Anti-SF antibodies Ⅱ and anti-sTfR antibodies Ⅱ were used as detection antibodies and goat antibodies coupled to Cy3 were used as antibodies Ⅲ.The detection conditions of the quantitative analysis method for simultaneous measurement of SF and sTfR with protein microarray were optimized and evaluated.The protein microarray was compared with commercially available traditional tests with 26 serum samples.Results By comparison experiment,mouse monoclonal antibodies were chosen as the probes and contact printing was chosen as the printing method.The concentrations of SF and sTfR probes were 0.5mg/mL and 0.5 mg/mL respectively,while those of SF and sTfR detection antibodies were 5 μg/mL and 0.36 μg/mL respectively.Intra- and inter-assay variability was between 3.26％ and 18.38％ for all tests.The regression coefficients comparing protein microarray with traditional test assays were better than 0.81 for SF and sTfR.Conclusion The present study has established a protein microarray method for combined measurement of SF and sTfR.