Novel coronavirus antibody quality control product and preparation method thereof
A quality control product and antibody technology, applied in the field of immunodetection, can solve the problems of uneven quality of new coronavirus antibody quality control reagents, high coincidence rate of test results, and complicated sample collection process, so as to achieve uniform product structure and increase freeze-drying. Efficiency, effect of reducing freeze-drying time
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[0020] The preparation method of the new coronavirus antibody quality control product of an embodiment of the present invention comprises the following steps:
[0021] The new coronavirus antibody is mixed with the freeze-drying protection solution to obtain the antibody solution.
[0022] The antibody solution was subjected to pre-freezing treatment, main drying treatment and final drying treatment in sequence to obtain the new coronavirus antibody quality control product.
[0023] Among them, the condition parameters of the pre-freezing treatment are as follows: partition temperature -5℃~-15℃ for 0.5 hours to 1 hour, partition temperature -45℃~-55℃ for 2.5 hours to 3.5 hours, partition temperature -15 ℃~-25℃ for 1.5 hours to 2.5 hours, partition temperature -45℃~-55℃ for 3.5 hours to 4.5 hours; the condition parameters of the main drying treatment are: partition temperature -45℃~-55℃, vacuum Temperature of 0.35mbar~0.45mbar for 25min~35min, partition temperature of -45℃~-55...
Embodiment 1
[0041] Example 1 Antibody Screening Preparation
[0042] Equal amounts of recombinant 2019-nCoV protein and Freund's adjuvant were mixed and fully emulsified, and three Balb / c mice were injected with 25 μg per mouse, and immunized three times with an interval of 15 days.
[0043] Cell fusion, selective culture, and screening: 3 days before fusion, mice were intraperitoneally injected with 25 μg of immune protein, and the spleen cells of immunized mice were taken out, and 2.0×10 7 2.0×10 SP2 / 0 myeloma cells 8 Splenocytes of 1 immunized Balb / c mouse were mixed, centrifuged, discarded supernatant, slightly oscillated and mixed, in 37°C water bath, 1 mL of 50% PEG-1500 aqueous solution was added dropwise within 90 seconds, Then add 20 mL of 1640 medium dropwise, centrifuge, discard the supernatant, wash once more, centrifuge, discard the supernatant to obtain hybridoma cells. Hybridoma cells were selected on a 96-well cell culture plate using HAT selection medium, and the total ...
Embodiment 2
[0064] Embodiment 2 quality control product preparation
[0065]Screening of raw materials: First, screen positive materials from different sources. The sources of positive materials include chimeric antibodies prepared above, conventional monoclonal antibodies, and conventional polyclonal antibodies. The three antibodies are 25 times and 50 times the same with the same deimmunoglobulin serum. , 100 times, 200 times, 400 times, 800 times, 1600 times serial dilution, after dilution, test its concentration with a kit (chemiluminescence method), the results show that the above-mentioned chimeric antibody > conventional monoclonal antibody > conventional polyclonal antibody, concentration The data are shown in Table 6. After confirming the activity of the raw materials, the three raw materials were prepared with immunoglobulin-free serum to prepare negative and positive target concentration samples, and their uniformity was tested. The results showed that the homogeneity results o...
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