Method to separate and purify recombinant human lactoferrin from rice seeds

摘要

A chromatography method for separating and purifying recombinant human lactoferrin from transgenic rice seeds, sequentially comprises the following steps of: 1) extracting recombinant human lactoferrin from transgenic rice seeds, to obtain the crude extract containing lactoferrin recombinant human; 2) subjecting the crude extract containing recombinant human lactoferrin to cation exchange chromatography to perform primary purification, to obtain an elution fraction containing recombinant human lactoferrin; where: cation exchange chromatography is performed on a CM sephorose FF weak cation exchange resin, the chromatography step comprises: 2A) equilibrating the chromatography column with 5-15 times the volume of buffer column containing Tris 10-25 mM, 10-25 mM NaAC, 0-250 mM NaCl, pH 6.0-7.5, at a flow rate of 50-200 cm / h; 2B) use the crude extract from step 1) as a loading sample, where the sample has a conductance of 2-25 ms / cm, and a pH of 6.0-7.5; elute the impurities with an impurity wash buffer, containing the impurity wash buffer NaH2PO4 3-10 mM, Na2HPO4 3-10 mM, NaCl 200-300 mM, pH 6.5-8.0, and the flow rate being elution of 50-200 cm / h; 2C) elute the sample with a wash buffer containing 3-10 mM NaH2PO4, 3-10 mM Na2HPO4, 400-650 mM NaCl, pH 6.5-8.0 at a flow rate of 50-200 cm / h, for obtain an elution fraction; 3) collect the elution fraction from step 2), to obtain the solution containing recombinant human lactoferrin.