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Device for detecting sugar chain abnormal IgA kidney disease and kit using said device

A kit and sugar chain technology, applied in the biological field, can solve the problems of no progress, not accepted by patients, and expensive surgery.

Inactive Publication Date: 2007-10-10
BEIJING HOTGEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Renal biopsy is an invasive operation with certain risks, can cause complications such as hematuria, and is expensive, so it is often not accepted by patients
[0014] There is a clinical need for a kit that can quickly and efficiently detect abnormal sugar chain IgA, but there has been no progress
The reason for this difficulty is that the difference between normal IgA and abnormal sugar chain IgA1 only exists in sugar chains, that is, there are only slight qualitative changes but no obvious quantitative changes, and the previous methods cannot effectively distinguish between the two

Method used

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  • Device for detecting sugar chain abnormal IgA kidney disease and kit using said device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Production of an enzyme-linked immunosorbent assay kit for detecting abnormal sugar chain IgA in IgA nephropathy:

[0086] The composition of the kit (96 persons) includes:

[0087] IgA nephropathy sugar chain abnormal IgA affinity adsorption centrifuge tube 96 copies

[0088] 1 vial of IgA nephropathy negative and positive controls;

[0089] Goat anti-human IgA polyclonal antibody-coated plate (96 wells) 1 piece;

[0090] 1 bottle of horseradish peroxidase (HRP)-labeled mouse anti-human IgA1 monoclonal antibody, 6ml / bottle;

[0091] 1 bottle of IgA standard;

[0092] 1 bottle each of substrate solution A and chromogenic solution B, 5ml / bottle;

[0093] 1 bottle of reaction termination solution, 5ml / bottle;

[0094] Abnormal sugar chain IgA washing buffer 25ml / bottle

[0095] Abnormal sugar chain IgA eluent 15ml / bottle

[0096] Concentrated washing solution (20 times concentrated solution, 20×) 20ml / bottle

[0097] The specific operation is as follows...

Embodiment 2

[0115] Example 2: Production of a chemiluminescent detection kit for detecting abnormal sugar chain IgA in IgA nephropathy

[0116] The composition of the kit (96 persons) includes:

[0117] IgA nephropathy sugar chain abnormal IgA affinity adsorption centrifuge tube 96 copies

[0118] IgA nephropathy negative, 1 bottle of positive control;

[0119] Goat anti-human IgA polyclonal antibody-coated plate (96 wells) 1 piece;

[0120] 1 bottle of horseradish peroxidase (HRP)-labeled mouse anti-human IgA1 monoclonal antibody, 6ml / bottle;

[0121] 1 bottle of IgA standard

[0122] 1 bottle each of substrate solution A and chromogenic solution B, 5ml / bottle;

[0123] Abnormal sugar chain IgA washing buffer 25ml / bottle

[0124] Abnormal sugar chain IgA eluent 15ml / bottle

[0125] Concentrated washing solution (20 times concentrated solution, 20×) 20ml / bottle

[0126] The specific operation is as follows:

[0127] 1. Preparation of IgA nephropathy-negative and positive controls:...

Embodiment 3

[0143] Example 3: Preparation and use of prepacked spin columns

[0144] 1. The prepacked spin column is composed of a spin column and a packing medium. The spin column is composed of an upper centrifuge tube and a lower collection tube. The two are set together to form a spin column. Auxiliary reagents include sugar chain abnormal IgA cleaning solution and sugar chain Abnormal IgA eluate.

[0145] 2. Preparation of spin column packing materials: Sepharose4B activated by CNBr from Pharmacia and HAA from Sigma were used for coupling according to the following steps:

[0146] (1) Soak 1.5g Sepharose 4B in 1mmol / L HCl until it swells, transfer it into a sand core funnel, and wash it with 300mL 1mmol / L HCl for about 30min;

[0147] (2) Weigh 50mg of HAA, dissolve in 7.5mL coupling buffer (0.1mmol / L NaHCO3, pH8.3, 0.5mol / L NaCl), combine with washed Sepharose 4B, and turn it upside down in a 10mL test tube with stopper Mix well (room temperature, 2h);

[0148] (3) Wash away uncoup...

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Abstract

A detection device of sugar chain abnormal IgA kidney disease is a preset centrifugal column for separating sugar chain abnormal ZgA of IgA kidney disease prepared as packing affine media of HAA in top separation tube and erecting filter cloth at bottom of top separation tube, packing buffer solution in bottom collection tube, combining HAA with sugar chain abnormal IgA to obtain sugar chain abnormal IgA combined with HAA by centrifugal elution. The detection kit applying said preset centrifugal column is also disclosed.

Description

technical field [0001] The invention relates to a prepacked spin column for separating abnormal sugar chain IgA of IgA nephropathy and a kit using the device, which belongs to the field of biotechnology and is used for diagnosing the occurrence of IgA nephropathy. Background technique [0002] IgA nephropathy (IgAN) is a group of chronic glomerular diseases caused by multiple etiologies with the same immunopathological features. Clinically, about 40%-45% of the patients showed gross or microscopic hematuria, 35%-40% showed microscopic hematuria with proteinuria, and the rest showed nephrotic syndrome and renal failure. IgA nephropathy is a common glomerular disease in the world, and its prevalence varies greatly in different continents, different countries or in different regions of a country, such as Japan and Singapore in Asia, the incidence of IgA nephropathy accounts for the primary 50% of glomerular diseases, while the low incidence of Indians in the western United Sta...

Claims

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Application Information

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IPC IPC(8): G01N1/34G01N33/53G01N33/577G01N21/76
Inventor 林长青
Owner BEIJING HOTGEN BIOTECH CO LTD
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