Objective: To optimize the conditions of protein microarray assay for soluble transferrin receptor (sTfR).
Methods: A microarrayer was used for printing anti-sTfR as antibody I on each protein microarray; anti-sTfR antibody was used as detection antibody II and goat antibody coupled to Cy3 was used as antibody III. The standard sTfR was detected by double antibody sandwich technique.
Results: Mouse monoclon sTfR antibody was chosen as a probe and contact printing as the printing method. A better homogenicity was appeared after pre-printing for 40 spots. The concentration of sTfR probes was 0.25 mg/ml. The concentration of sTfR detection antibody was 0.18 microg/ml. 3% skim milk powder and goat anti rabbit antibody made by GE was chosen as blocking buffer and secondary antibody. The lowest detection limits and the biological detection limits of sTfR were 0.253 ng/ml and 0.78 ng/ml respectively. The best fitting models and standard curve were established for sTfR (r = 0.99699).
Conclusion: Conditions of a quantitative analysis system for measurement of sTfR with protein microarray were optimized.