Method for improving detection specificity of immuno-reagent and application of method

Abstract

The invention provides a method for immuno-detection. According to the method, pepsin is utilized first to treat a murine monoclonal antibody, and then the treated murine monoclonal antibody is utilized to perform immuno-detection. The immuno-detection method provided by the invention significantly reduces false positive results of the immuno-detection. Moreover, by using a diluent provided by the invention, the detection value of a false positive sample can be further reduced, and the biological stability of a maker of the treated murine monoclonal antibody, preserved at a working concentration, can be improved. The diluent is a 4-(2-hydroxyerhyl)piperazine-1-ethanesulfonic acid buffer solution, and contains casein, magnesium sulfate and a compound enzyme stabilizer DPD, and the pH value is adjusted to 7.0-8.0 by using a sodium hydroxide solution.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents. Specifically, the present invention relates to a method for improving the detection specificity of an immune reagent and its application. Background technique [0002] In vitro diagnostic reagents refer to reagents, kits, calibrators (substances), quality control products (substances), etc. used for in vitro detection of human samples (various body fluids, cells, tissue samples, etc.). The doctor's prescription has a very important prompting effect. [0003] Among in vitro diagnostic reagents, the immunological method based on double antibody sandwich technology is a very commonly used method for detecting antigens, and is widely used in enzyme-linked immunoassay, colloidal gold method, and chemiluminescence reagents. Among the raw materials of various double-antibody sandwich detection reagents, mouse-derived monoclonal antibodies (hereinafter referred to as mouse monoclonal antibod...

AI Technical Summary

Problems solved by technology

At present, in addition to mouse monoclonal antibodies, there are also rabbit monoclonal antibodies, but the preparation process of rabbit monoclonal antibodies is not mature, and only a few companies have the production capacity of this antibody, and the price is very expensive

[0008] 3. Adding a higher concentration of irrelevant free mouse monoclonal antibody to the reagent reduces the detection value through the method of competitive inhibition

First of all, the composition of commercial blocking substances is not fully disclosed, and it is not known whether the confidential components in the reagent will affect the test results of other types of special samples

Second, commercial blocking substances are only effective for some special samples

Furthermore, such blocking substances are often expensive and used in large quantities, which is not conducive to the cost control of reagent manufacturers in large-scale production.

[0010] To sum up, in the immunoassay reagents with double monoclonal antibody sandwich method to detect antigen as the reaction system, if the sample contains HAMA or RF, it is very easy to produce false positive results, which brings certain risks to clinical diagnosis and doctor's prescription. troubled

At present, the commonly used solutions in the industry either have poor performance, which affects the performance of diagnostic reagents; or the process is immature and cannot be stably and repeatedly produced; or the cost is relatively high, which is a heavy burden on the enterprise.

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