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E. coli host cell proteins （HCP） constitute a diverse and functionally intricate group of macromolecules within the Escherichia coli cell. The host proteins of E. coli play crucial roles in various cellular processes. These proteins are involved in fundamental metabolic pathways such as glycolysis, the tricarboxylic acid cycle, and amino acid biosynthesis, which are essential for the survival and growth of the bacterium. Recombinant expression in E. coli is a simple and cost-effective method for producing certain proteins. These proteins, often used as therapeutic agents, must be highly purified. The manufacturing process may introduce contamination by HCPs from E. coli, which can reduce efficacy and cause adverse reactions. Therefore, minimizing HCP impurities is crucial. This simple, sensitive, and fast OneStep ELISA is a powerful method for optimizing purification processes, process control, routine quality control, and product release testing.