Novel detection method and kit for synchronously detecting concentration of free kappa light chain and free lambda light chain

A technology of simultaneous detection and kits, applied in the field of immunoassays, which can solve problems such as complex and cumbersome detection processes, and unsatisfactory sensitivity and precision

Inactive Publication Date: 2012-10-17
SHANGHAI HUJING BIO TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Therefore, the technical problem to be solved in the present invention is to detect the free kappa light chain and the free lambda light chain respectively in the existing method. Large and produce hook effect and other shortcomings, provide a method and kit for simultaneous detection of free kappa light chain and free lambda light chain, the detection method and kit have high sensitivity, strong specificity, good stability and simple operation , fast, can greatly improve the efficiency of diagnosis

Method used

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  • Novel detection method and kit for synchronously detecting concentration of free kappa light chain and free lambda light chain
  • Novel detection method and kit for synchronously detecting concentration of free kappa light chain and free lambda light chain
  • Novel detection method and kit for synchronously detecting concentration of free kappa light chain and free lambda light chain

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Experimental program
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Effect test

Embodiment 1

[0045] The preparation of embodiment 1 kit

[0046] The reagents in each box are sufficient for 96 measurements, and the materials in the box are as follows:

[0047] (1) 1×96-well plate (8 strips×12 wells, which can be split into single wells) is coated with rabbit anti-human IgG H&L polyclonal antibody.

[0048] (2) 6× free kappa light chain / free lambda light chain mixed standard, lyophilized product, including 6 concentrations: 0mg / L, 5mg / L, 20mg / L, 100mg / L, 200mg / L, 500mg / L . Dissolve each bottle with 1ml of distilled water before use.

[0049] (3) 1× Europium-labeled antibody 1 freeze-dried product, dissolved in 0.5ml distilled water before use.

[0050] (4) 1 × samarium-labeled antibody 2 lyophilized product, dissolved in 0.5ml distilled water before use.

[0051] (5) 1× buffer solution, 30ml.

[0052] (6) 1×washing solution, 30ml, dilute 1:25 with distilled water when used.

[0053] (7) 1× enhancement solution, 15ml.

[0054] The following are the specific ingred...

Embodiment 2

[0073] The preparation of embodiment 2 kit

[0074] Materials in the kit: except for the 96-well plate coated with rabbit anti-human IgG H&L polyclonal antibody, the others are the same as in Example 1.

[0075] The preparation method of each material in the kit is as follows:

[0076] (1) Eu 3+ Preparation of labeled rabbit anti-human free kappa light chain polyclonal antibody

[0077] Refer to Eu 3+ Labeling kit (manufacturer PerkerElmer) manual operation. Specifically: Take 1 mg of rabbit anti-human free kappa light chain polyclonal antibody and dissolve it in 0.25mol / L NaHCO 3 Add 0.2mg Eu to 200μl 3+ -N 2 -[p-isocyanate-benzyl]-diethylenetriaminetetraacetic acid (Eu 3+ -DTTA) and mix thoroughly, and react at 4°C for 24 hours. The reaction solution was chromatographed on a Sephadex G-50 column (1×20 cm), monitored by A280 to collect protein peaks, combined peak tubes, and determined the protein content. At the same time with PerkinElmer Eu 3+ Determination of Eu ...

Embodiment 3

[0081] The preparation of embodiment 3 kits

[0082] The reagents in each box are sufficient for 48 measurements, and the materials in the box are as follows:

[0083] (1) 1×48-well plate (4 strips×12 wells, which can be split into single wells) is coated with mouse anti-human free kappa light chain monoclonal antibody.

[0084] (2) 6× free kappa light chain and free lambda light chain mixed standard, lyophilized product, including 6 concentrations: 0mg / L, 5mg / L, 20mg / L, 100mg / L, 200mg / L, 500mg / L . Dissolve each bottle with 1ml of distilled water before use.

[0085] (3) 1× europium-labeled antibody freeze-dried product, dissolved in 0.5ml distilled water before use.

[0086] (4) 1× samarium-labeled antibody freeze-dried product, dissolved in 0.5ml distilled water before use.

[0087] (5) 1× buffer solution, 30ml.

[0088] (6) 1×washing solution, 30ml, dilute 1:25 with distilled water when used.

[0089] (7) 1× enhancement solution, 15ml.

[0090] The preparation method...

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Abstract

The invention discloses a time-resolved fluoroimmunoassay (TRFIA) kit and a detection method thereof for synchronously detecting the concentration of a free kappa light chain and a free lambda light chain. The kit comprises: 1) an antibody synchronously aiming at a heavy chain and a light chain of a human antibody, 2) an anti-free kappa light chain antibody labelled by lanthanide, 3) an anti-free lambda light chain antibody labelled by another lanthanide, 4) a standard substance of the free kappa light chain and a standard substance of the free lambda light chain, 5) a buffer, 6) a washing liquid, and 7) an enhancement solution. According to the invention, the antibody synchronously aiming at the heavy chain and the light chain of the human antibody is used as a capture antibody, other two antibodies respectively aiming at the free kappa light chain and the free lambda light chain are used as the labelled antibodies, double labeling TRFIA detection principles are used, thus the novel detection method for synchronously detecting the concentration of the free kappa light chain and the free lambda light chain is established. The method provided by the invention has the advantages of high sensitivity, strong specificity and good stability, and can realize high degrees of automation.

Description

technical field [0001] The invention belongs to the field of immune detection, in particular to a time-resolved fluorescent immunoassay method and a kit for synchronously detecting free kappa light chains and free lambda light chains. Background technique [0002] Immunoglobulin (Ig) light chains are divided into two types, kappa (kappa) and lambda (lambda), each Ig molecule has only one type of light chain, and the ratio of human kappa (kappa) and lambda (lambda) is 6:4. Light chains are small molecular proteins that can freely pass through the glomerular basement membrane, and are reabsorbed back into the blood circulation in the renal tubules, so only a small amount of light chains exist in normal human urine. When metabolic disorders and multiple myeloma occur, a large number of free light chains will appear in the blood and be excreted in the urine, which is Bence-Jones protein. [0003] Significant basic and clinical studies have shown that serum free light chain det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 盛世乐王金凤
Owner SHANGHAI HUJING BIO TECH CO LTD
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