Method for high-throughput detection of EB virus infection efficiency/antibody blocking EB virus infection efficiency
An Epstein-Barr virus, high-throughput technology, applied in the field of biomedicine, can solve the problems of inability to meet high-throughput detection, 96-well plate reading detection, time-consuming and other problems, achieve high-throughput detection methods, reduce time, The effect of simple detection methods
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Embodiment 1
[0043] Embodiment 1 High-content instrument detects the infection efficiency of ten batches of Epstein-Barr virus
[0044]1. The virus production cell line is cultivated to 90% cell confluency, and then the chemical inducer phorbol ester (final concentration 20ng / ml) and sodium butyrate (final concentration 2.5mM) are used to jointly induce the virus production cell line to produce virus, The cell culture supernatant was harvested 72 hours after induction, the virus was purified and concentrated by high-speed centrifugation, and the virus was subpackaged and stored at -80 degrees for later use.
[0045] 2. Ten batches of CNE2-EBV virus production;
[0046] 3. Dilute each batch of virus liquid into 8 gradients according to a 2-fold gradient, then mix the diluted virus liquid with the cells and incubate in a 37-degree carbon dioxide incubator for 2 hours;
[0047] 4. After 2 hours, the virus supernatant was removed by centrifugation, and then the cells were resuspended with com...
Embodiment 2
[0058] Example 2 Using a high-content detection instrument to detect the neutralizing effect of neutralizing antibody 72A1 in blocking Epstein-Barr virus infection
[0059] 1. The virus production cell line is cultivated to 90% cell confluency, and then the chemical inducer phorbol ester (final concentration 20ng / ml) and sodium butyrate (final concentration 2.5mM) are used to jointly induce the virus production cell line to produce virus, The cell culture supernatant was harvested 72 hours after induction, the virus was purified and concentrated by high-speed centrifugation, and the virus was subpackaged and stored at -80 degrees for later use.
[0060] 2. The production of CNE2-EBV virus is as described above; and three batches of Epstein-Barr virus with infection efficiencies of 10%, 20%, and 40% are selected as blocking objects;
[0061] 3. The neutralizing antibody 72A1 was diluted 5 times and co-diluted 8 gradients according to the initial concentration of 5mg / ml for bloc...
Embodiment 3
[0071] Example 3 Using flow cytometry to detect the neutralizing effect of neutralizing antibody 72A1 in blocking Epstein-Barr virus infection, and analyzing the correlation between the two detection methods
[0072] 1. The virus production cell line is cultivated to 90% cell confluency, and then the chemical inducer phorbol ester (final concentration 20ng / ml) and sodium butyrate (final concentration 2.5mM) are used to jointly induce the virus production cell line to produce virus, The cell culture supernatant was harvested 72 hours after induction, the virus was purified and concentrated by high-speed centrifugation, and the virus was subpackaged and stored at -80 degrees for later use.
[0073] 2. The production of CNE2-EBV virus is as described above; and three batches of Epstein-Barr virus with infection efficiencies of 10%, 20%, and 40% are selected as blocking objects;
[0074] 3. The neutralizing antibody 72A1 was diluted 5 times and co-diluted 8 gradients according to ...
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