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Method for recovering conjugate from mother liquor of antibody coupling medicine

A technology of antibody-conjugated drugs and conjugates, applied in the field of medicine

Active Publication Date: 2022-04-05
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is no relevant report on the separation and purification of the conjugate from the mother solution of antibody-conjugated drugs

Method used

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  • Method for recovering conjugate from mother liquor of antibody coupling medicine
  • Method for recovering conjugate from mother liquor of antibody coupling medicine
  • Method for recovering conjugate from mother liquor of antibody coupling medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] This example provides a method for recovering the conjugate from the mother liquor of the antibody-drug conjugate, and the specific steps are as follows:

[0068] Pretreatment: at 25-35°C, weigh 30g type A non-polar macroporous adsorption resin, and pack it into a column by wet method (resin column volume is 50cm 3 ), then add 100mL absolute ethanol to clean the resin column, control the cleaning speed to be 2BV / h, then clean the resin column with 300mL deionized water to no ethanol smell, and set aside;

[0069] Sample loading: At 25-35°C, measure 100mL of the mother solution of the above-mentioned antibody-conjugated drug, pass through the resin column after the above pretreatment at a flow rate of 2BV / h, open the knob at the lower end of the glass chromatography column, and let the mother solution flow out slowly. During the sampling process, add a nitrogen balloon to the upper end of the resin column for protection;

[0070] Impurity removal and desorption: After l...

Embodiment 2

[0073] This example provides a method for recovering the conjugate from the mother liquor of the antibody-drug conjugate, and the specific steps are as follows:

[0074] Pretreatment: at 15-25°C, weigh 120g type A non-polar macroporous adsorption resin, and pack it into a column by wet method (resin column volume is 200cm 3 ), then add 400mL absolute ethanol to clean the resin column, the control cleaning speed is 2BV / h, then use 800mL deionized water to clean the resin column to the smell of ethanol, for subsequent use;

[0075] Sample loading: At 15-25°C, measure 400mL of the mother solution of the above-mentioned antibody-conjugated drug, pass through the resin column after the above pretreatment at a flow rate of 2BV / h, open the knob at the lower end of the glass chromatography column, let the mother solution flow out slowly, and During the sample loading process, add a nitrogen balloon to the upper end of the resin column for protection;

[0076] Impurity removal and des...

Embodiment 3

[0079] This example provides a method for recovering the conjugate from the mother liquor of the antibody-drug conjugate, and the specific steps are as follows:

[0080] Pretreatment: take by weighing 300g type A nonpolar macroporous adsorption resin, and use wet packing column (resin column volume is 500cm 3 ), then add 1000mL absolute ethanol to clean the resin column, control the cleaning speed to be 2BV / h, then clean the resin column with 1800mL deionized water to no ethanol smell, and set aside;

[0081] Sample loading: at 25-35°C, measure 1000mL of the mother solution of the above-mentioned antibody-conjugated drug, pass through the resin column after the above pretreatment at a flow rate of 1BV / h, open the knob at the lower end of the glass chromatography column, let the mother solution flow out slowly, and During the sample loading process, add a nitrogen balloon to the upper end of the resin column for protection;

[0082] Impurity removal and desorption: After loadi...

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Abstract

The invention discloses a method for recovering a conjugate from mother liquor of an antibody-coupled drug, which comprises the following steps: loading the mother liquor of the antibody-coupled drug to non-polar macroporous adsorption resin, and adsorbing the conjugate; removing impurities from the non-polar macroporous adsorption resin adsorbed with the conjugate by using an impurity removal solution, desorbing by using an eluent, collecting the eluent containing the conjugate, and concentrating, solventing out and crystallizing to obtain the conjugate, wherein the non-polar macroporous adsorption resin is a styrene series or a styrene-divinylbenzene series. According to the method, the non-polar macroporous adsorption resin with specific filler is adopted, and the subsequent elution crystallization step is combined, so that the yield and the purity of the conjugate recovered from the mother liquor of the antibody coupling medicine can be remarkably improved, the yield can reach 59% or above, the purity can reach 95% or above, the utilization rate of raw materials is greatly improved, and the method is suitable for industrial production. The preparation cost of the antibody coupling medicine is reduced.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for recovering conjugates from the mother liquor of antibody-conjugated drugs. Background technique [0002] Antibody-Drug Conjugate (ADC) is composed of three parts: antibody, cytotoxic drug, and conjugated chain, that is, the cytotoxic drug and the antibody with tumor targeting are coupled to form a whole through the conjugated chain. It is a new generation of tumor-targeted drugs to make full use of the specificity and targeting of antibodies to tumor cell-associated antigens, and to deliver cytotoxic drugs to tumor lesions. [0003] Since the cytotoxic drugs in the ADC require high toxicity and affinity, and at the same time have low sensitivity to the efflux mediated by the multidrug resistance protein (MDR1), there are fewer applicable cytotoxic drugs, which makes antibody conjugation Drug development is difficult, and there are fewer antibody-conjuga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/02C07K1/22C07K1/30C07K1/36
Inventor 尚振华李婷焦德琛
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY