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Pre-mounted eccentric column for detecting liver cancer alpha-fetoprotein heteroplasmon and reagent kid containing same

An alpha-fetoprotein and spin column technology, which is applied in the field of medical products, can solve the problems of complicated operation and no domestic reagent supply, and achieve the effects of accurate results, fast detection and simple method.

Active Publication Date: 2007-02-28
BEIJING HOTGEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The whole experimental process is complicated. For a long time, only a few clinical laboratories in China can measure AFP heterogeneity, and a few cities can meet the requirements of clinical determination of AFP heterogeneity, and there is no domestic reagent supply.

Method used

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  • Pre-mounted eccentric column for detecting liver cancer alpha-fetoprotein heteroplasmon and reagent kid containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Production of an enzyme-linked immunosorbent assay kit for detecting alpha-fetoprotein heterogeneity in liver cancer:

[0076] The composition of the kit (96 persons) includes:

[0077] 1 bottle of AFP-L3 negative and positive controls;

[0078] AFP monoclonal antibody coated plate (96 wells) 1 piece;

[0079] 1 bottle of horseradish peroxidase (HRP)-labeled AFP monoclonal antibody, 6ml / bottle;

[0080] 1 bottle of AFP standard;

[0081] 1 bottle each of substrate solution A and chromogenic solution B, 5ml / bottle;

[0082] 1 bottle of reaction termination solution, 5ml / bottle;

[0083] 1 bottle of washing buffer (20×concentrated), 30ml / bottle.

[0084] The specific operation is as follows:

[0085] 1. Prepare AFP-negative and positive controls:

[0086] 1) Serum collection: Obtain the serum of healthy normal people and ascites of liver cancer patients from the hospital or blood bank, and store it at -70°C for later use;

[0087] 2) Packing:

[0088] ...

Embodiment 2

[0125] Example 2: Production of a chemiluminescent detection kit for detecting alpha-fetoprotein heterogeneity in liver cancer

[0126] The kit (48 servings) consists of:

[0127] One bottle of AFP-L3 negative and one positive control;

[0128] AFP monoclonal antibody coated plate (96 wells) 1 piece;

[0129] 1 bottle of horseradish peroxidase (HRP)-labeled AFP monoclonal antibody, 6ml / bottle;

[0130] 1 bottle of AFP standard

[0131] 1 bottle each of substrate solution A and chromogenic solution B, 5ml / bottle;

[0132] Wash buffer (20X concentrated) 1 bottle, 30ml / bottle.

[0133] The specific operation is as follows:

[0134] 1. Prepare AFP-negative and positive controls:

[0135] 1) Serum collection: Obtain normal human serum and ascites of liver cancer patients from hospitals or blood banks, and store them at -70°C for later use;

[0136] 2) Packing:

[0137] AFP-L3 positive control substance: Ascites from patients with liver cancer positive, put into 1.5ml leppen...

Embodiment 3

[0173] Example 3: Preparation and use of prepacked spin columns

[0174] 1. The prepacked spin column is composed of a spin column and a filling 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.

[0175] 2. Preparation of spin column packing materials: Sepharose4B activated by CNBr from Pharmacia and LCA from Sigma were used for coupling according to the following steps:

[0176] (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;

[0177] (2) Weigh 2mg of LCA, dissolve in 7.5mL coupling buffer (0.1mmol / L NaHCO3, pH8.3, 0.5mol / L NaCl), combine with washed Sepharose 4B, mix upside down with a stoppered 10mL test tube Uniform (room temperature, 2h);

[0178] (3) Wash off uncoupled LCA with 10 mL of coupling buffer. After measuring the LCA I content in the washing liquid, the calculated coupli...

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Abstract

The invention relates to a preformed eccentric post for checking the a-fetoprotein alloplasm of liver cancer, wherein said eccentric post is formed by upper separating tube and lower collecting tube; the separating tube contains affinity medium of coupled small lentils agglutinin (LCA); the bottom of separating tube is one filter cloth; the lower collecting tube contains buffer solution; said LCA can combine the a-fetoprotein alloplasm sugar chain. The invention also provides a relative chemical lighting testing agent box of said preformed eccentric post and an enzyme couple immunity quantitative testing agent box with preformed eccentric post, and relative preparation and application. Said agent box can quickly and simply test the content of a-fetoprotein alloplasm, to diagnose liver cancer.

Description

technical field [0001] The present invention relates to a prepacked spin column for detecting liver cancer alpha-fetoprotein heterogeneity, and also relates to a chemiluminescent detection kit containing the prepacked spin column and an enzyme-linked immunosorbent assay containing the prepacked spin column Quantitative detection kits and preparation methods of these kits belong to the technical field of medical products and are used for early diagnosis of liver cancer. Background technique [0002] Hepatocellular carcinoma (hepatocellular carcinoma, HCC) is the fourth most common malignant tumor in the world. About 250,000 patients die of hepatocellular carcinoma every year. The pathogenesis of hepatocellular carcinoma is still unclear. The degree of death is relatively high, and the mortality rate ranks third among malignant tumors of the gastrointestinal tract. Therefore, early detection and early treatment are the key to improving the survival rate of patients. Factors i...

Claims

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Application Information

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IPC IPC(8): G01N1/34G01N33/577G01N33/543G01N21/76
CPCB01L2300/047B01L3/502G01N2333/471B01L3/5021G01N1/405G01N33/57438B01L3/50825
Inventor 林长青
Owner BEIJING HOTGEN BIOTECH CO LTD
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