Method and kit for detecting human serum hybrid light chain antibody

A kit and human detection technology are applied in the field of clinical laboratory science to achieve the effects of convenient use, strong specificity and simple operation

Inactive Publication Date: 2015-08-26
卫生部北京医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No studies have reported the existence of hybrid light chain antibody IgG4 in patients with autoimmune pancreatitis

Method used

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  • Method and kit for detecting human serum hybrid light chain antibody
  • Method and kit for detecting human serum hybrid light chain antibody
  • Method and kit for detecting human serum hybrid light chain antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1: Kit for detecting hybrid light chain antibodies in human serum

[0056] 1. Kit composition

[0057] 1. Anti-human lambda light chain antibody coated microwell plate;

[0058] 2. Anti-human kappa light chain antibody labeled with horseradish peroxidase: Abcam Company;

[0059] 3. Washing solution (PBST): PBS solution containing 0.05% Tween-20; (add 500 μL Tween-20 to 1L PBS, mix well)

[0060] 4. Diluent: PBST containing 20% ​​(v / v) calf serum;

[0061] 5. Substrate solution: 3′,3′,5′,5′,-tetramethylbenzidine (TMB);

[0062] 6. Standard reference serum: 20 cases of healthy people with the same volume of mixed serum, the specified concentration of hybrid antibody is 1AU / ml;

[0063] 7. Negative control solution: PBST of 20% calf serum by volume;

[0064] 8. Stop solution: 0.5M sulfuric acid.

[0065] 2. Conditions and methods for preparing anti-human lambda light chain-coated microwell plates:

[0066] A: Microplate: Nunc, 96-well plate, Denmark;

[006...

Embodiment 2

[0072] Example 2: Detection of Human Serum Hybrid Light Chain Antibody

[0073] 1. Materials

[0074] 1. the test kit described in embodiment 1

[0075] 2. Serum: Serum from 34 patients with autoimmune pancreatitis, 60 patients with pancreatic cancer, 42 disease controls (including 23 cases of acute pancreatitis and 19 cases of chronic non-autoimmune pancreatitis), 62 serum samples from healthy people . Stored at -80°C until analysis.

[0076] 2. Method

[0077] 1. Detection of heterozygous light chain antibodies by double-antibody sandwich method

[0078] (1) All materials and reagents were equilibrated to room temperature before measurement;

[0079] (2) Dilute the standard serum: Use a sample gun to draw 1 μL of the standard serum provided by the kit and add it to 1 mL of the diluent to prepare a 1AU / uL stock solution, then draw 500 uL and add it to 1 ml of the diluent, and perform 6 consecutive times of 2 times Gradual dilution to prepare standard serum with concentr...

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Abstract

The invention relates to a method and a kit for detecting a human serum hybrid light chain antibody, and belongs to the field of clinical laboratory medicine. The kit consists of the following components: (1) an anti-human lambda light chain antibody enveloped porous plate; (2) an immunolabeling anti-human kappa light chain antibody; (3) a scrubbing solution (PBST); (4) a diluent; (5) a substrate solution : 3',3',5',5',-tetramethyl benzidine; (6) standard reference serum; (7) a negative control solution; (8) a stop solution. The method and the kit are used for an antibody-sandwich enzyme linked immunosorbent assay to detect the human serum hybrid light chain antibody, and the method is simple and easy, and can serve as a new method for performing differential diagnosis on autoimmune pancreatitis and pancreatic cancers.

Description

technical field [0001] The invention relates to a kit for detecting hybrid light chain antibodies in human serum, which belongs to the field of clinical laboratory science. Background technique [0002] In the past ten years, many experiments have confirmed that IgG4 molecules can form bispecific antibodies through "half-molecule exchange" (Fab arm exchange) in vivo and in vitro. This process includes the re- The exchange between the light chain pairs and the regeneration of the disulfide bonds in the hinge region can generate an immunoglobulin with two different antigen-binding domains at the same time. Serine at position 228 in the hinge region of the antibody and arginine at position 409 in the CH3 region as well as serum reduced glutathione (GSH) are essential for this exchange between antibody molecules. There are two types of immunoglobulin light chains in the human body, namely kappa and lambda. The genes encoding the two light chains are located on the chromosome at...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57438G01N33/57488G01N33/6893G01N2800/067
Inventor 李金明郝明巨
Owner 卫生部北京医院
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