	US 12,352,759 B2
	Assay method of antibody
Rajeshwari Patel, Ahmedabad (IN); and Praveen Kumar Anidil Kizhakinagath, Ahmedabad (IN)
Assigned to Kashiv BioSciences, LLC, Piscataway, NJ (US)
Filed by Kashiv BioSciences, LLC, Piscataway, NJ (US)
Filed on Jul. 11, 2024, as Appl. No. 18/769,708.
Application 18/769,708 is a continuation of application No. PCT/IB2023/056773, filed on Jun. 29, 2023.
Claims priority of application No. 202221037354 (IN), filed on Jun. 29, 2022.
Prior Publication US 2025/0003984 A1, Jan. 2, 2025
Int. Cl. G01N 33/53 (2006.01); C12Q 1/40 (2006.01); G01N 33/68 (2006.01)

CPC G01N 33/6854 (2013.01) [C12Q 1/40 (2013.01); G01N 2333/70535 (2013.01); G01N 2333/765 (2013.01); G01N 2333/924 (2013.01)]

19 Claims

1. An assay method comprising:

a) seeding RBL 2H3 hFCεRIa cells in an assay plate with a MEM media to form a cell suspension, wherein the RBL 2H3 hFCεRIa cells comprise an hFCεRIa receptor with an IgE or an NP IgE and an anti-IgE antibody;

b) optionally incubating the cell suspension with CO2,

c) sensitising and neutralizing the RBL 2H3 cells;

d) transferring the sensitized and neutralized RBL 2H3 cells obtained from step (c) to a pre-seeded assay plate;

e) incubating the pre-seeded assay plate;

f) washing the pre-seeded assay plate;

g) mixing an antigen into the pre-seeded assay plate and incubating the pre-seeded assay plate to obtain a supernatant;

h) transferring the supernatant obtained from the incubation in step (g) from the pre-seeded assay plate into a test assay plate;

i) incorporating a substrate into the test assay plate and incubating the test assay plate; and

j) measuring bioactivity of the anti-IgE antibody.