Immunogenicity analysis kit for double-tolerance anti-IgE monoclonal antibody drug as well as using method and application of immunogenicity analysis kit
A technology of immunogenicity and analytical reagents, applied in the field of biological analysis and drug immunogenicity analysis, it can solve the problems of weak positive, affecting drug resistance and detection sensitivity, and false negative.
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Embodiment 1
[0089] A Bridge ECL method was constructed using biotin-labeled drugs and ruthenium-labeled drugs. Considering the need to overcome the interference of free IgE in the serum, (another) anti-IgE monoclonal antibody (ligelizumab, ligelizumab) was used to coat the 96 microwell enzyme plate (ELISA plate). First, the positive polyclonal antibody (ADA, positive anti-drug antibody) of anti-IgE monoclonal antibody prepared in the serum sample and the serum sample without adding ADA were mixed with anti-IgE monoclonal antibody (Drug, Omalizumab) according to the concentration values described in Table 1. Monoclonal antibody) was acidified for the first time after dilution according to the minimum dilution factor (MRD, 10 times). The volume ratio of the acidifying solution to the sample solution to be acidified is 1:9. Neutralize the sample solution after the first acidification. The volume ratio of the neutralizing solution and the sample solution after the first acidification is 1...
Embodiment 2
[0094] It is basically the same as in Example 1, the only difference is that the IgE antibody is replaced by the IgE receptor in the ELISA plate. That is to say, in Example 1, another anti-IgE monoclonal antibody was used to affinity capture and remove IgE, and in Example 2, the IgE receptor (ligand anti-drug target, Human Fc epsilon RI alpha Protein) was coated on the enzyme label plate (ELISA plate), the remaining implementation steps are the same as in Example 1.
[0095] The detection analysis result of table 2 embodiment 2
[0096]
[0097] Compared with Example 1, using another anti-IgE antibody to capture IgE, Example 2 uses IgE receptor affinity to capture IgE in blood samples, which is more conducive to improving the drug tolerance of the analysis method. It can be seen from Table 2 that the signal intensity of Example 2 is generally increased, especially blood samples containing different concentrations of ADA can almost all tolerate 50 μg / mL of the drug. Compar...
Embodiment 3
[0100]The technical scheme of double enzyme plate combined with direct ECL is used to improve drug resistance and sensitivity. Specifically: use (another) monoclonal antibody of anti-IgE monoclonal antibody to coat on the microtiter plate (ELISA plate 1), and take another microtiter plate (ELISA plate 2) at the same time, and coat it on this microtiter plate Anti-IgE monoclonal antibody (Drug, Omalizumab). First, the positive polyclonal antibody (ADA, positive anti-drug antibody) of anti-IgE monoclonal antibody prepared in the serum sample and the negative serum sample without adding ADA were mixed with anti-IgE monoclonal antibody (Drug, Oma Zhuzumab) mixture was acidified for the first time after dilution according to the minimum dilution factor (MRD, 10 times). The volume ratio of the sample to be acidified and the acidified solution is 1:9. Neutralize the sample solution after the first acidification. The volume ratio of the sample solution after the first acidification...
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