We present a new low-cost technology to fabricate poly(pyrrole) microwires (PPy-μWs) by micro contact printing (μCP) and controlled chemical polymerization (μCP-CCP) using three different techniques (“grafting printing”, “addressed printing”, and “direct printing”. These developed techniques provide a platform to regulate PPy wire size and shape using N-(3-trimethoxysilyl-propyl) pyrrole (Py-silane) for the “addressed printing” and “direct printing” techniques, respectively. The technique uses a silicon mold to produce micropatterned polydimethylsiloxane (PDMS) stamps. The micropatterned PDMS stamp was employed to print PPy-μWs with an improvement observed from “direct printing” > “addressed printing” > “grafting printing”. Therefore, for “direct printing”, PPy-μWs were printed on: poly(ethylene terephthalate) (PETE), cyclic olefin copolymer (COC), poly(ethylene 2,6-naphthalate (PEN), and polyimide (PI) surfaces. Here, we have studied the printing of wires and the surface morphology to understand the characteristics of PPy patterns by optical microscopy, scanning electrode microscopy (SEM), and atomic force microscopy (AFM). The functionality of the PPy-μW matrix was also analyzed for immune detection. Here, fluorescent imaging was used to certify the detection of recombinant human interleukin-10 (rh IL-10). PPy-μWs were functionalized using gluteraldehyde (GA) for the specific immobilization of anti-human interleukin-10 monoclonal antibodies (anti-IL-10 mAb's). The immunosensor present a sensitivity of 0.026 (pg/mL⁻¹) to the respective rh IL-10 in a linear range of 1–50 pg/mL and a limit of detection at 0.36 pg/L. Finally the PPy-μW matrix was successfully tested for cytokine bioassays using electrochemical impedance spectroscopy (EIS) and presents outstanding properties as a matrix for biosensing.