SU1586740A1 - Method of producing sorbent for affine chromatography - Google Patents

Abstract

This invention relates to biospecific sorbents and permits an increase in sorption capacity and a reduction in the duration of the process for the preparation of a sorbent. Cotton fibers are treated with an aqueous solution of sodium periodate to an oxidation level of 5-10%, then the carrier is treated with 5-20% albumin solution in physiological solution at PH 8-10 for 6 hours. The carrier is washed with water and treated with 2-5% hormone solution in 70% dioxane in the presence of carbodiimide. The product is washed with dioxane. 80% aqueous ethanol and water. Get the sorbent with a hormone content of 15-25 mg / g. 2 tab.

Description

This invention relates to biospecific sorbents and can be used to isolate pure protein receptors and hormone antibodies.

The aim of the invention is to increase the sorption capacity and reduce the duration of the process for obtaining a sorbent T 3.

Example 1. The south of the cotton fibers is treated with an aqueous solution of the period- I ratio of iodic acid U, U 7035 g per 1 g of fibers at pH 4.5, module 5 and room temperature for 24 hours. Fiber is obtained with an oxidation state of 5 X. The fiber obtained is treated with a 5% solution of albumin in buffered saline at pH 8, module 5 for 6 hours at 20 ° C.

The fiber is washed with water for 1 hour at module 10 at 20 ° C and carried out

immobilization of hydrocortisone by treating it with a 2% solution in 70% aqueous dioxane in the presence of 50 mg of 1-methyl-3- (3-dimethylaminopropyl) -carbodimimide. Processing is continued for 4 hours and another 50 mg of carbodiimide is added. Treatment module 5. Time 14 hours at 20 ° C.

The target product was washed with dioxane for 1 hour at a 10 h module with 80% aqueous ethanol at a 51 h module and WATER at a 10-15 module. Washing quality control is carried out spectrophotometrically. The target product contains 50 mg / g of albumin and 15 mg / g of hydrocortisone hemisuccinate (Table 1).

Example 2. 10 g of cotton fibers are oxidized as in example 1, but with an iodic acid ratio of 0.99849 g per 1 g of fiber A fiber with an oxidation state of 7% is obtained. The resulting fiber is treated

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10% albumin solution in buffered saline at pH 9, module 5 for 6 hours at 20 ° C. The fiber is washed with water and the hydrocortisone is immobilized by treating it with a 3% solution of 70% aqueous dioxane under the conditions of Example 1. The washing is carried out according to Example 1. The target product contains 100 mg of albumin per 1 g of carrier and 20 mg of hydrocortisone 1 g of carrier.

Example 3. Oxidation of cotton fibers was carried out as in Example 1, but with a ratio of iodic acid of 0.1407 g per 1 g of fiber. A fiber with an oxidation state of 10% is obtained. The fiber obtained is treated with a 20% albumin solution in buffered saline at pH 10, module 5 for 6 hours at 20 ° C. The fiber is washed with water and treated with 5% hydrocortisone B with 70% aqueous dioxane under conditions of x Example 1. The rinsing is carried out as in Example 1, the desired product contains 200 mg of albumin and 25 ml of hormone per 1.g carrier.

Example 4. The south of the cotton fibers is treated as in Example 1 to obtain a fiber containing 50 mg / g of albumin. Instead of hydrocortisone hemisuccinate, a 2% solution of estradiol hemisuccinate is added to the albumin-containing fiber and immobilized in Example 1 for hydrocortisone hemsuccinate. The target product contains 50 mg / g of albumin and 16 mg / g of estradiol hemisuccinate.

Sorption properties are given in table 2.

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Example 5. The south of the cotton fibers is treated as in Example 2. But instead of hydrocortisone hemisuccinate, the same amount of estradiol hemisuccinate is added. The washing is carried out according to Example 1. The target product contains 100 mg of albumin per 1 g of carrier and 22 mg of estisdiol hemisuccinate per 1 g of carrier (Table 2).

Example 6. Oxidation of 10 g of cotton fiber and immobilization of albumin on it is carried out according to Example 3. When treating with a hormone, the same amount of estradiol hemisuccinate (5% solution in 70% dioxane under the conditions of example 1) is added instead of hydrocortisone. The washing is carried out according to Example 1. The target product contains 200 mg of albumin and 27 mg of the hormone per gram of carrier (Table 2).

Data on the sorption capacity of sorbents obtained by the proposed and known methods are presented in table. 1 and 2.

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From tab. 1 and 2, it is clear that the proposed method makes it possible to obtain a sorbent containing 10 times more immobilized hormone, with a sorption capacity of 220–370 times more, than 20 sMoro obtained from the sorbent by means of a known sssoyu. The process time is reduced from 73 to 40 hours, and the semi-adsorbent does not require special storage conditions and can be stored at atmospheric pressure and room temperature.

Claims (1)

25 Formula of Invention A method for producing an affinity chromatography sorbent, comprising sequentially treating a textile carrier containing aldehyde groups with aqueous 30 albumin solution. And a hormone solution in 70% aqueous dioxane in the presence of carbodiimide, followed by washing the resulting sorbent, characterized in that, in order to increase the sorption 35 capacity and reduce the duration of the process, as a textile carrier use aldehyde-containing cellulosic material with an oxidation state of 5-10%, the processing is carried out 5-20% 40 solution of albumin in physiological solution of prN 8-U to an albumin content of 50–200 mg / g of carrier and then 2–5% solution of the hormone to contain 15–25 mg / g of carrier and the product is sequentially washed with dioxane, 80% ethanol and water. Table 1 table 2