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Purification of papain by immobilized metal affinity chromatography (IMAC) on chelating carboxymethyl cellulose

机译:螯合羧甲基纤维素上的固定金属亲和色谱法(IMAC)纯化木瓜蛋白酶

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Chelating carboxymethyl cellulose was prepared in bead form by immobilizing iminodiacetic acid on carboxymethyl cellulose which was earlier crosslinked and activated by epichlorohydrin. The prepared matrix was used to purify papain by a factor of 2.6 from commercial papain, and by a factor of 4 from papaya latex by batch adsorption and immobilized metal affinity chromatography respectively. Purification factors obtained were equal in batch mode and double in column mode, to purifications obtained on Chelating Sepharose~(~R) Fast Flow. Flow rates up to 38 ml/cm~2 h were easily possible on the prepared chelating carboxymethyl cellulose.
机译:通过将亚氨基二乙酸固定在较早被表氯醇交联和活化的羧甲基纤维素上,以珠形式制备螯合的羧甲基纤维素。所制备的基质分别通过分批吸附法和固定化金属亲和色谱法分别从商品木瓜蛋白酶中纯化出2.6倍的木瓜蛋白酶和从木瓜乳胶中纯化得到的四倍。获得的纯化因子在批处理模式下是相等的,在柱模式下是两倍,与在螯合Sepharose快速流中获得的纯化因子相同。在制备的螯合羧甲基纤维素上,流速很容易达到38 ml / cm〜2 h。

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