The development of a robust surface functionalization method is indispensable in controlling the efficiency, sensitivity, and stability of a detection system. Polystyrene (PS) has been used as a support material in various biomedical fields. Here, we report various strategies of polystyrene surface functionalization using siloxane derivative, divinyl sulfone, cyanogen bromide, and carbonyl diimidazole for the immobilization of biological recognition elements (peptide developed to detect ochratoxin A) for a binding assay with ochratoxin A (OTA). Our objective is to develop future detection systems that would use polystyrene cuvettes such as immobilization support of biological recognition elements. The goal of this article is to demonstrate the proof of concept of this immobilization support. The results obtained reveal the successful modification of polystyrene surfaces with the coupling agents. Furthermore, the immobilization of biological recognition elements, for the OTA binding assay with horseradish peroxidase conjugated to ochratoxin A (OTA-HRP) also confirms that the characteristics of the functionalized peptide immobilized on polystyrene retains its ability to bind to its ligand. The presented strategies on the functionalization of polystyrene surfaces will offer alternatives to the possibilities of immobilizing biomolecules with excellent order- forming monolayers, due to their robust surface chemistries and validate a proof of concept for the development of highly efficient, sensitive, and stable future biosensors for food or water pollution monitoring.