METHOD AND KIT FOR DETERMINING- ANTIBODY SENSITIVITY AND CLONE CELL STRAIN

• US 20140371090A1
• Filed: 06/30/2014
• Published: 12/18/2014
• Est. Priority Date: 12/31/2011
• Status: Active Grant

• Current Assignee: Shenzhen Mindray Bio-Medical Electronics Co., Ltd. (Excelsior Union Ltd.)
• Original Assignee: Shenzhen Mindray Bio-Medical Electronics Co., Ltd. (Excelsior Union Ltd.)
• Inventors: Lei, Ting, Zhao, Yumei, Dong, Lifang
• Granted Patent: US 9,632,086 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2812 against CD4
• CPC Class Codes: C07K 16/289 against CD45
• CPC Class Codes: G01N 2333/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: G01N 2333/70514 CD4
• CPC Class Codes: G01N 2333/70589 CD45
• CPC Class Codes: G01N 2333/70596 Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/554 the carrier being a biological cell or cell fragment, e.g. bacteria, yeast cells
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

A method and kit for determining antibody sensitivity and quality of a clone cell stain. The method comprises: obtaining a solid-phase carrier, cells and an antibody; adsorbing the antibody on the solid-phase carrier; incubating the cells and the antibody; preserving cells bound with the antibody; and dyeing and counting the cells bound with the antibody, so as to determine the antibody sensitivity or the quality of the clone cell stain based on the cell count. The kit comprises components used in the method.

Claims

1. A method for determining antibody sensitivity, comprising:

obtaining a solid-phase carrier, cells and an antibody;

adsorbing the antibody on the solid-phase carrier;

incubating the cells and the antibody;

preserving cells bound with the antibody;

dyeing and counting the cells bound with the antibody; and

determining the antibody sensitivity based on the cell count.

3. The method according to claim 1, wherein

determining the antibody sensitivity based on the cell count, the higher the cell count, the higher the antibody sensitivity;

4. The method according to claim 1, wherein said cells are cells from body fluid of human or animal, or primary or generation cells from cell culturing.

5. The method according to claim 4, wherein said cells from body fluid of human or animal are selected from red blood cells, leukocytes or platelets, preferred cells is selected from sub-populations cells of leukocyte.

6. The method according to claim 1, wherein said antibody is selected from monoclonal antibody.

7. The method according to claim 1, wherein said solid-phase carrier is selected from cellulose, cross-linked dextran or polystyrene.

8. The method according to claim 1, wherein said cells are prepared as a cell suspension having a concentration ranging 5-500 cells/μ

l.

9. The method according to claim 1, wherein the cells and the antibody are incubated at 20°

C.-25°

C. for one hour.

10. The method according to claim 1, wherein:

before adsorbing the antibody on the solid-phase carrier, the antibody is mixed with a coating buffer having a pH value ranging 6-8.

11. The method according to claim 10, wherein said coating buffer has an ionic concentration ranging 0.01-1 mol/L.

12. The method according to claim 1, further comprising:

dyeing living cells bound with the antibody using a dye solution, wherein the dye in the dye solution is selected from at least one of methyl chloride, Switzerland Giemsa solution, sand yellow and hematoxylin.

13. The method according to claim 12, wherein the methyl chloride is dissolved in water with a concentration ranging 0.05-5%.

14. The method according to claim 1, further comprising:

fixing the cells bound with the antibody before dyeing the cells.

15. The method according to claim 1, further comprising:

performing a positive and/or negative control, which comprises;

providing an antibody with known sensitivity as a positive and/or negative antibody;

adsorbing the positive and/or negative antibody on the solid-phase carrier;

incubating the cells and the positive and/or negative antibody;

preserving cells bound with the positive and/or negative antibody;

dyeing and counting the cells bound with the positive and/or negative antibody.

17. A kit for antibody or clone cell strain determination using method of claims 1, said kit comprising:

a solid-phase carrier, which is used to adsorb an antibody which is to be tested or secreted by a clone cell strain to be tested;

a coating buffer, in which the antibody is adsorbed on the solid-phase carrier;

a cell washing solution, which is used to remove cells unbound with the antibody; and

a dye solution, which is used to dye the cells bound with the antibody.

18. The kit according to claim 17, said kit further comprising:

a cell fixing solution, which is used to fix the cells bound with the antibody.

20. The method according to claim 1, wherein said antibody is selected from monoclonal antibody against antigen expressed on a cell surface.

21. The method according to claim 1, wherein said antibody is selected from monoclonal antibody against surface antigen of lymphocyte.

2. (canceled)

16. (canceled)

19. A method for determining antibody specificity, said method comprising:

obtaining a solid-phase carrier, cells and an antibody, the cells including at least two different known types cells;

adsorbing the antibody on the solid-phase carrier;

incubating the cells and the antibody adsorbed on the solid-phase carrier;

preserving cells bound with the antibody;

dyeing and counting the cells bound with the antibody; and

determining the antibody specificity based on the cell count of the two different known type cells.

22. A method for determining clone cell strain, comprising:

providing a solid-phase carrier, cells and an antibody secreted by a clone cell strain;

adsorbing the antibody secreted by the clone cell strain on the solid-phase carrier;

incubating the cells and the antibody;

preserving cells bound with the antibody;

dyeing and counting the cells bound with the antibody; and

determining the clone cell strain based on the cell count.

23. A method according to claim 22, wherein

determining the clone cell strain based on the cell count, the higher the cell count, the better quality of the clone cell strain.