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We developed a new kind of suspension array for multiplexed immunoassays using silica colloidal crystal beads (SCCBs) as coding carriers. The monodisperse and size-controlled SCCBs were fabricated by a microfluidic device. Calcination was employed to improve the mechanical stability and lower the fluorescent background of the SCCBs. Immobilization of protein molecules on the surface of the SCCBs through chemical bonds was studied, and the modification condition was optimized to increase the detection sensitivity. Results indicated that the SCCBs as supports were more sensitive (0.92 ng/mL IgG) than the glass beads (27 ng/mL IgG) and the planar carriers (140 ng/mL IgG). A multiplex immunoassay showed the flexibility and feasibility of SCCBs array in clinical applications.