Objective: This study aimed to optimize the process conditions for the large-scale production of milk-derived casein glycomacropeptide （CGMP） and explore its hemagglutination inhibition （HI） ability against influenza virus. Methods: Using anion exchange resin as the separation medium and sialic acid content the activity detection index of the product,based on the preliminary small and medium-sized tests, the large-scale production of CGMP from whey powder was investigated from five process conditions: sample volume, loading flow, eluent concentration, eluent volume, and resin service cycle. Meanwhile, a hemagglutination inhibition test was used to verify the inhibitory activity of the product CGMP against the influenza virus. Results: The optimized technical route of CGMP product is as follows: loading volume 3BV, loading flow 0.8 L/min, elution volume 1BV, eluent concentration 0.6 mol/L, and suitable use cycle of resin 4 times. Under these conditions, the volume of whey powder solution was 108 L （2 160 g） and the resin was 36 L, and 21.183 g of CGMP product was obtained, with a yield of 0.981 g/100 g, the product purity of 89.032%, the recovery rate of 51.972% and the sialic acid content of 21.503 g/100 g. The minimum inhibitory concentration of CGMP products on influenza virus A （H1N1） and influenza virus B is 1.000 0 and 1.250 0 mg/mL, respectively. Conclusion: The purity and sialic acid content of CGMP produced by the optimized technical route were better than those of commercial CGMP, and had an obvious inhibitory effect on the influenza virus.