This application note describes how to achieve accurate comparability assessments using Biacore™ T200 and Amersham™ WB system in the development of a biosimilar. This is exemplified in this study by the use of these systems in the high-throughput process development (HTPD) of a biosimilar molecule, interferon α-2a (IFNα-2a)*. Biacore T200 was extensively used starting from the refolding step, through filtration procedures, followed by three chromatographic purification steps: capture, intermediate purification, and polishing. Two types of surface plasmon resonance (SPR) experiment made it possible to: (i) assess the comparability to the originator molecule; (ii) find the best refolding conditions; (iii) select the most appropriate chromatographic media (resins); and (iv) optimize the binding and elution steps. Amersham WB system was used to analyze the purity and confirm accurately the identity of the IFNα-2a biosimilar with fluorescence multiplexing detection.