This paper presents a simple and reliable one-touch type multi-immunosensing lab-on-a-chip (LOC) detecting antibodies as multi-disease markers using electrochemical method suitable for a portable point-of-care system (POCS). The multi-stacked LOC consists of a PDMS space layer for liquids loading, a PDMS valve layer with 50 im in height for the membrane, a PDMS channel layer for the fluid paths, and a glass layer for multi electrodes. For the disposable immunoassay which needs sequential flow control of sample and buffer liquids according to the designed strategies, reliable and easy-controlled on-chip operation mechanisms without any electric power are necessary. The driving forces of sequential liquids transfer are the capillary attraction force and the pneumatic pressure generated by air bladder push. These passive fluid transport mechanisms are suitable for single-use LOC module. Prior to the application of detection of the antibody as a disease marker, the model experiments were performed with anti-DNP antibody and anti-biotin antibody as target analytes. The flow test results demonstrate that we can control the fluid flow easily by using the capillary stop valve and the PDMS check valves. By the model tests, we confirmed that the proposed LOC is easily applicable to the bioanalytic immunosensors using bioelectrocatalysis.