The new recombinant protein purification protocol combines lysis and purification into a single step. The condensed workflow can be completed in about 45 minutes, reducing processing time by 75% compared to traditional techniques, while delivering high yield and purity. The method uses a specialized lysis buffer in combination with magnetic affinity capture beads, which together eliminate the need to clarify lysates by centrifugation. As a result, the workflow can be automated using a high-throughput particle processor, further reducing sample variability. The one-step lysis and purification protocol has been validated specifically for PureProteome™ Nickel Magnetic Beads, optimized for the capture of histidine-tagged proteins, and BugBuster® Master Mix, a lysis reagent that allows for nonmechanical extraction of soluble protein from bacterial cells. This reagent combines detergent-based lysis with enzymatic agents, Benzonase® nuclease, and rLysozyme™ enzyme, in a ready-to-use formulation.