Optimization of purification protocol of biomimetic affinity material for marine metalloprotease MP

Objective: To establish an efficient method for biomimetic affinity purification of marine metalloprotease MP.Methods: Using the biomimetic affinity materials to optimize the purification conditions by response surface optimization method, such as the single factor experiment, Plackett-Burman experiment design, Box-Behnken experiment.And the purity of purified samples by SDS-PAGE and HPLC was analyzed. Results: The purification conditions were optimized by single factor experiment and the response surface design. The study indicated that the best purification conditions of the boronic acid gel were: When enzyme concentration was 100 mg/m L, the optimal loading buffer was glycine-sodium hydroxide of p H8.6 and the sample rate was 1 m L/min. The elution buffer was disodium hydrogen phosphate-citric acid of p H5.2, and elution rate was2 m L/min, the elution buffer added to mannitol of 75 mmol/L, the purification efficiency was enhanced 11 times and the purity above 98.8%.