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用阳离子交换树脂CM-52分离纯化类人胶原蛋白II(HumanCollagen-likeBioproteinII,HCBII)。通过实验,分别确定了静态吸附和柱层析吸附的操作条件,即静态吸附:在pH4.0、NaCl0.15mol/L、进料浓度7g/L,处理量17.26ml/g树脂的条件下进行吸附,吸附时间为60min;柱层析:在pH4.0、NaCl0.15mol/L、进料浓度5g/L、流速5ml/min的条件下进行吸附。然后上柱,在pH4.0、NaCl0.30mol/L下进行脱附。实验表明,采用静态吸附的方式吸附HCBII,然后上柱洗脱,HCBII的吸附量可达到48.6mg/g树脂,回收率83.8%,操作时间短,分辨率高,最终纯化的HCBII达电泳纯,分子量为97kD。
Human collogen-like Bioprotein II (HCBII) was isolated and purified by cation exchange resin CM-52. Through the experiments, the operating conditions of static adsorption and column chromatography were determined respectively, that is, the static adsorption was carried out under the conditions of pH4.0, NaCl0.15mol / L, feed concentration7g / L and treatment volume17.26ml / g resin Adsorption, adsorption time was 60min; Column chromatography: pH4.0, NaCl0.15mol / L, the feed concentration of 5g / L, the flow rate of 5ml / min under the conditions of adsorption. Then the column was desorbed at pH 4.0, NaCl0.30 mol / L. The experimental results showed that HCBII was adsorbed by static adsorption and then eluted on the column. The adsorption capacity of HCBII reached 48.6mg / g, the recovery rate was 83.8%, the operation time was short and the resolution was high. The final purified HCBII reached electrophoretic purity, The molecular weight is 97kD.