Patterned silver nanoparticle (NP)-poly[2-(methacryloyloxy)ethyl] trimethyl ammonium chloride (AgNP-polyMETAC) composites were prepared by electrochemical lithography, surface-initiated atom-transfer radical polymerization (SI-ATRP) and NP growth inside the polymer brushes. For this purpose, polymer brushes of poly[2-(methacryloyloxy)ethyl] trimethyl ammonium chloride (polyMETAC) were utilized as strong electrolyte brush system. These were introduced in form of patterned polymer brushes to create pH-responsive surface enhanced Raman scattering SERS substrates. It is well-known that the charges of strong polyelectrolyte chains are usually insensitive to pH changes, hence, rarely strong polyelectrolyte brushes have been utilized so far to study pH-responsive properties of such films. Here pH-insensitive polyMETAC brushes exhibit pH-sensitive properties and can be used as pH-responsive surfaces for SERS applications due to the embedding of AgNPs into the polymer brushes. When increasing the pH, the assembly of the AgNPs transfers from quasi two-dimensional (2D) aggregates, attaching mainly to the polymer surface, into a three-dimensional (3D) assembly, where the particles are penetrating into the brushes. These changes result in significant alterations of the SERS efficiency of the polymer brush composite. At pH 5, the enhancement of the Raman scattering approaches its maximum. The fabricated SERS substrates show a high sensitivity as well as good experimental reliability at different pH values. Moreover, electrochemical lithography was utilized to fabricate patterned SERS substrate, which allows an easy combination of multiple other functionalities in hierarchical structuring steps. In addition, the microstructure is in our studies beneficial because of a simplified and reliable characterization of the polymer brushes at defined sample areas. The introduction of the microstructured brush system is regarded moreover attractive for the development of high-throughput platforms for rapid, automated screening and analysis applications.