A preparation method of affinity chromatography medium for separating and purifying hexokinase
A technology for separation and purification of hexokinase, which is applied in the field of preparation of affinity chromatography media, and achieves the effects of low price, easy acquisition and good application prospects
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Embodiment 1
[0032] The invention provides a method for preparing an affinity chromatography medium for separating and purifying chitosanase, comprising the following steps:
[0033] S1. First, take 5 g of washed and drained agarose 6FF gel and place it in a 250 ml Erlenmeyer flask, add 50 ml of sodium periodate solution with a concentration of 0.01 g / mL, place it in a constant temperature shaking incubator, and control the temperature at 25 ℃, and the rotation speed was 150rpm to carry out the aldylation reaction for 1h, and the aldylation agarose 6FF gel was prepared;
[0034] S2. Take 1 g of the aldylated agarose gel prepared in step S1, put it in a sand core funnel, wash it with distilled water, drain it, transfer it to a 250 mL conical flask, add 50 mL of glucosamine solution with a concentration of 0.01 g / mL, Place in a constant temperature shaking incubator, control the temperature at 37°C, and carry out the coupling reaction at 150rpm for 12h;
[0035] S3, transfer the agarose gel...
Embodiment 2
[0047] The invention provides a method for preparing an affinity chromatography medium for separating and purifying hexokinase, comprising the following steps:
[0048] S1. First, take 2 g of washed and drained agarose 6FF gel, 1 g of agarose CL-6B gel, and 2 g of agarose 4B gel, put them into a 250 ml Erlenmeyer flask, and add 50 ml of high concentration of 0.03 g / mL The sodium iodate solution was placed in a constant temperature shaking incubator, the temperature was controlled at 25°C, and the rotation speed was 150rpm, and the aldylation reaction was carried out for 1 hour to prepare the aldylated agarose 6FF gel;
[0049] S2. Take 1 g of the aldylated agarose gel prepared in step S1, place it in a sand core funnel, wash it with distilled water, drain it, transfer it to a 250 mL Erlenmeyer flask, add 50 mL of glucosamine solution with a concentration of 1 g / mL, Place in a constant temperature shaking incubator, control the temperature at 37°C, and carry out the coupling re...
Embodiment 3
[0061] The invention provides a method for preparing an affinity chromatography medium for separating and purifying hexokinase, comprising the following steps:
[0062] S1. First, take 2 g of washed and drained agarose 6FF gel, 1 g of agarose CL-6B gel, and 2 g of agarose 4B gel, put them into a 250 ml Erlenmeyer flask, and add 50 ml of high concentration of 0.03 g / mL The sodium iodate solution was placed in a constant temperature shaking incubator, the temperature was controlled at 20°C, and the rotation speed was 250rpm, and the aldylation reaction was carried out for 2 hours to prepare the aldylated agarose 6FF gel;
[0063] S2. Take 1 g of the aldylated agarose gel prepared in step S1, place it in a sand core funnel, wash it with distilled water, drain it, transfer it to a 250 mL Erlenmeyer flask, add 50 mL of glucosamine solution with a concentration of 1 g / mL, Place in a constant temperature shaking incubator, control the temperature at 30°C, and carry out the coupling r...
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Abstract
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