Human cytomegalovirus cross-linked antibody and its application in immunoassay kit
A technology of human cytomegalovirus and cross-linking agent, which is applied in the direction of hybrid immunoglobulin, measuring devices, instruments, etc., can solve problems such as difficult standardization, difficult inactivation, and unstable quality, so as to reduce detection errors and improve production and preparation. Simple, Avoid Infectious Effects
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Embodiment 1
[0044] Utilize SPDP cross-linking agent (purchased from Thermo Scientific Company, product number 21857) to prepare the heterologous cross-linked product of mouse anti-HSV-II McAb and healthy human IgG not infected with HSV-II as HSV-II in patient's positive blood Antibody surrogate, the process is as follows:
[0045] (1) Dissolve 5 mg mouse anti-HSV-II McAb in cross-linking buffer (0.1M potassium phosphate, 0.1M NaCl, pH7.5), stir and add 50 μl SPDP cross-linking agent (3.2 mg / ml, dissolved in In absolute ethanol), after reacting at room temperature for 2 hours, put it into a dialysis bag, dialyze with reducing buffer (0.1M sodium acetate, 0.1M NaCl, pH4.5), and change the solution four times;
[0046] (2) Dissolve 5 mg of healthy human IgG uninfected with HSV-II in the cross-linking buffer, stir and add 50 μl SPDP cross-linking agent, react at room temperature for 2 hours, put it into a dialysis bag, and dialyze with the cross-linking buffer , change the liquid four times;...
Embodiment 2
[0053] Example 2 Experiments on substitution of mouse anti-HSV-II McAb and healthy non-HSV-II-infected human IgG heterologous cross-linked product, quality control stability and cryopreservation stability
[0054] Experiments were carried out using the mouse anti-HSV-II McAb prepared in Example 1 and the heterologous cross-linked product of healthy human IgG not infected with HSV-II as a substitute for the HSV-II antibody in the patient's positive blood.
[0055] (1) Preparation of quality control products. Prepare the heterologous cross-linked products in which the mass ratio of the tetrameric heterologous cross-linked antibody in the prepared heterologous cross-linked product is 50%, 70%, 90% and 100% in sequence as the patient Surrogates for HSV-II antibodies in positive blood. When the heterologous cross-linked antibody in the form of tetramer accounts for 50%, 70% or 90% of the mass ratio in the prepared heterologous cross-linked product, the remaining 50%, 30% or 10% he...
Embodiment 3
[0075] Example 3 Alternative experiment of mouse anti-HSV-1 McAb and healthy non-infected HSV-1 human IgG heterologous crosslinker
[0076] Using the SPDP cross-linker to prepare mouse anti-HSV-I McAb and healthy non-HSV-I-infected human IgG heterologous cross-linked product as a surrogate for HSV-I antibody in patient-positive blood, except for mouse anti-HSV -I McAb replaces mouse anti-HSV-II McAb, and the specific process is the same as in Example 1.
[0077] Experiments were carried out using the mouse anti-HSV-1 McAb prepared in Example 3 and the heterologous cross-linked product of healthy human IgG not infected with HSV-1 as a substitute for the HSV-1 antibody in the patient's positive blood. The heterologous cross-linked products in which the tetrameric heterologous cross-linked antibody accounts for 50%, 70%, 90% and 100% of the mass ratio in the prepared heterologous cross-linked product are sequentially prepared. When the heterologous cross-linked antibody in the f...
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