Method and kit for determining-antibody sensitivity and clone cell strain

• US 9,632,086 B2
• Filed: 06/30/2014
• Issued: 04/25/2017
• 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: Zhao, Yumei, Dong, Lifang, Lei, Ting
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US14/319,056
• Publication Number:
  US 20140371090A1
• Time in Patent Office: 1,030 Days
• 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 binding sensitivity, comprising:

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

adsorbing the antibody on the solid-phase carrier to form an adsorbed antibody;

incubating the cells and the adsorbed antibody;

washing the solid-phase to preserve cells bound with the adsorbed antibody;

dyeing the cells bound with the adsorbed antibody with a dye to obtain stained cells;

counting the stained cells one by one to obtain a number of the stained cells; and

determining the antibody binding sensitivity based on the number of the stained cells in comparison to a positive control and a negative control, wherein the higher the number of the stained cells as compared to the negative control, the better the antibody binding sensitivity.

2. The method according to claim 1, wherein counting the stained cells one by one is performed under a microscope.

3. The method according to claim 1, wherein said cells are cells from body fluid of human or animal;

or primary or generation cells that are obtained by cell culturing.

4. The method according to claim 3, wherein said cells from body fluid of human or animal are selected from the group consisting of red blood cells, leukocytes and platelets.

5. The method according to claim 4, wherein said cells are lymphocytes.

6. The method according to claim 1, wherein the antibody is a monoclonal antibody.

7. The method according to claim 1, wherein said solid-phase carrier is selected from the group consisting of cellulose, cross-linked dextran and polystyrene.

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

l.

9. The method according to claim 1, wherein the cells and the adsorbed 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 from 6-8.

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

12. The method according to claim 1,

wherein the dye is selected from the group consisting 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 from 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 binding 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;

washing the solid-phase to preserve cells bound with the positive and/or negative antibody;

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

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

17. The method according to claim 1, wherein said antibody is a monoclonal antibody against surface antigen of lymphocyte.