Mutation and aggregation of superoxide dismutase (SOD) are reported as one of the causes of amyotrophic lateral sclerosis (ALS). To detect SOD1 protein from the motor neuron, surface plasmon resonance (SPR) analysis was adopted due to its advantages in the in-situ biomolecular recognition by surface analysis without labeling. For the patterning of protein antigens at Au surface for use in SPR imaging experiments, microfluidic devices were fabricated with polydimethylsiloxane (PDMS) by replica molding method. They were designed for the solution to flow by capillary force only without using any additional pumping equipments or flow controllers. Performance of microfluidic devices was verified by the simple microfluidic experiments, and multiple protein-patterned sensor surfaces were constructed by using these microfluidic devices.