Patent ID: 11921114
Assignee: nan
Field: Measurement (Instruments)
Classification: CPC G  C | IPC C  G

Claim 8:
9. A process for performing microneutralization assays to quantify neutralizing antibodies to viruses, the process comprising:
coating a plate with a monolayer of Vero E6 cells;
inoculating the coated plate with a serum-included inoculum, the serum-included inoculum comprising:
serum from a patient, the serum being diluted; and
a virus, the virus being Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2);

incubating the inoculated plate for an initial incubating period of approximately one (˜1) hour at a temperature of ˜37±2° C. and a carbon-dioxide (CO2) content of ˜5±2% CO2; and
transferring the serum-included inoculum to the plate coated with the Vero E6 cells;

incubating further the plate with the fresh inoculation medium for a total incubating period of approximately forty-eight (˜48) hours at ˜37±2° C. and ˜5±2% CO2;
removing the inoculation medium after the total incubation period;
washing the plate with Hanks Buffered Salt Solution (HBSS) at approximately one-hundred-and-fifty microliters per well (˜150 μL/well) after removing the inoculation medium;
fixing the Vero E6 cells plate with approximately eighty percent (˜80%) acetone;
adding a primary antibody to the fixed plate, the primary antibody being selected from the group consisting of:
Mouse Species Coronavirus (COVID-19, MERS, and SARS-CoV NP) Antibody, Catalog Number HM1056 (“E3”);
E3 in combination with Mouse Species Coronavirus (COVID-19, MERS, and SARS-CoV NP) Antibody, Catalog Number HM1057 (“E1”); and
Mouse Species Anti-SARS-Cov-2 NP mAb, clone 4B21, Catalog Number CABT-CS027 (“C4”) in combination with Mouse Species Anti-SARS-Cov-2 NP mAb, clone 4B21, Catalog Number CABT-CS026 (“C5”);

incubating the plate with the added primary antibody;
washing the plate with the added primary antibody;
adding a secondary antibody to the plate;
incubating the plate with the added secondary antibody;
washing the plate with the added secondary antibody;
applying 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) solution;
applying a stop solution to the washed plate;
reading the optical density of the plate; and
determining whether the optical density is greater than approximately 0.7.