Operation method for improving drug tolerance in drug-resistant antibody analysis

Abstract

The invention discloses an operation method for improving drug tolerance in drug-resistant antibody analysis. The operation method comprises the following specific steps: acidifying a sample; carrying out SA magnetic bead cleaning and sealing; incubating the biotin labeled medicine; enriching the magnetic beads; acidifying the magnetic beads; and neutralizing and incubating. According to the operation method for improving drug tolerance in drug-resistant antibody analysis, the sample is treated through magnetic bead enrichment, the grabbing and enrichment efficiency of ADA is greatly improved, therapeutic protein products in the sample are effectively removed, the therapeutic protein products are prevented from being combined with ADA again, and the complexity of later detection is reduced. The ADA enriched by the magnetic beads can be dissociated through acidolysis and then can be subjected to later detection. Compared with a traditional method, the drug tolerance concentration of the method is improved by more than thousands of times.

Description

technical field [0001] The invention relates to the technical field of immunogenicity in biological analysis, in particular to an operation method for improving drug tolerance in anti-drug antibody analysis. Background technique [0002] In recent years, with the aging population and the continuous increase in the incidence of cancer, health issues have received widespread attention. Therefore, the biomedical industry has developed rapidly. More and more biological drugs have entered the stage of clinical trial research. In the long and complicated growth process, the failure of every link in the middle may lead to the termination of the project. There are many cases where the project fails when it is only one step away from the drug application for marketing. Among them, the drug safety caused by immunogenicity is the leading cause. One of the main reasons for failure; [0003] Immunogenicity is defined as the phenomenon that a therapeutic protein product produces an immun...

AI Technical Summary

Problems solved by technology

[0004] The detection method for immunogenicity currently mainly uses microplate reader, MSD and other means, mainly for the detection of immunogenic anti-drug antibody (ADA), but the problem lies in the detection of many ADA The method has the problem of poor drug tolerance, that is, the therapeutic protein product (the drug itself) or its endogenous target in the serum may interfere with the sensitivity of the method, which is especially obvious for drugs with long half-lives (such as antibody drugs), In the ADA detection method, the anti-interference ability of therapeutic protein products is evaluated, which is called drug tolerance. We know that anti-drug antibody detection is mainly to evaluate the production of therapeutic protein products against the immunogen after entering the human body. The ADA is aimed at the detection of ADA, not the drug itself, so the difficulty is that most of the subjects’ samples are treated with therapeutic protein products, and the ADA in the samples is a complex combined with therapeutic protein products , rather than a single form of ADA, so the sample must be pretreated before detection, the original combined complex is separated by acid hydrolysis, and then the ADA to be detected is obtained through the competition method, and finally the separated ADA is detected;

[0005] The main difficulty of the detection method for the study of ADA is to solve drug tolerance. The detection method currently developed is mainly to dissociate the originally bound complex through acid hydrolysis, and add a competitive reagent in an alkaline environment (usually the researched therapeutic protein product) after incubation for ADA detection to achieve the purpose of competing for ADA, but because the competition reagent is a solid phase and the concentration is not high, the competition ability is far lower than the binding efficiency of free drug and ADA; there are still a lot of acid in the sample after acid hydrolysis Therefore, the effect of this method on improving drug tolerance is not significant and has certain limitations

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