HLA complement-dependent cytotoxcity antibody detection method using ELIA as basis and its kit

A kit and antibody technology, applied in the field of CDC effect evaluation, can solve problems such as insufficient satisfaction, and achieve the effect of simple and easy to master method, low cost and high sensitivity

Inactive Publication Date: 2003-09-24
PEL FREEZ BIOTECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the above-mentioned methods for measuring HLA antibodies commonly used in the world cannot fully meet the above requirements.

Method used

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  • HLA complement-dependent cytotoxcity antibody detection method using ELIA as basis and its kit
  • HLA complement-dependent cytotoxcity antibody detection method using ELIA as basis and its kit
  • HLA complement-dependent cytotoxcity antibody detection method using ELIA as basis and its kit

Examples

Experimental program
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Effect test

example 4 HLA-I or / and HLA-IIELISA approach ( 1 ( or )HLA-I or / and HLA-II。( 2 example 3 ( 3 、( 4 ( 3 1 ； or 1 。；， and 2-3， 1 。( 4 example 3 ( 8 、( 9 、( 10 。 example 5 HLA CFAbs approach (CFAbs-PRA)( 1 PRA preparation ：HLA， 1 (0.01-3×106/)；CELISA； or HLA or 。( 2 PRA 1 1 ， and or ； example 3 ( 4 。( 3 or HLA；；；。( 4 HRP-C1qAb or HRP-C3Ab()， 1 ()。( 5 1 HRP，OD。( 6 ：ODCFAbs；PRA 100 ；：CFAbs÷PRA×100＝(PRA％) example ：CFAbs25；PRA50；PRA％50％。 example 6 HLA-I and HLA-IICFAbs( 1 HLA-I or / and HLA-II or ， example ，HLA-I or / and IICFAbs。( 2 HLA-ICFAbsHLA-I。( 3 HLA-I or II＞95％HLA。 example 7

[0032] Or the total lgG on the surface of B lymphocytes can be measured to achieve simultaneous determination of CFABS and Non-CFAbs. Example 3: ELISA method for HLA Class I / IICFAbs (1) Purified HLA Class I or / and Class II antigens are coated with a microtiter plate at 37°C for 1-4 hours or 4°C overnight. Aspirate and discard the package. By liquid. (2) Block the microplate with a buffer (blocking solution) containing a certain amount of non-specific protein (such as BSA, skimmed milk powder, calf serum, etc.), with the same time and temperature as step (1), and then aspirate the remaining blocking liquid. (3) Add 50 μl each of negative control serum, positive control serum, background buffer and test serum into the corresponding microplates (also can be serum diluted with buffer). (4) Incubate the above-mentioned microplate at 37°C or 4°C for 30 minutes to 3 hours or 4°C overnight. (5) Aspirate and discard the reaction solution, and wash each well of the microplate with a deterge...

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Abstract

According to the complement-dependent cytotoxicity (CDC) reaction principle in the immunology said invention creates an in-vitro enzyme-linked immunoreactino method for assaying HLA complement fixingantibodies (CFAbs) and its kit. The reaction system is formed from solid-phase HLA antigen or target cell with HLA antigen and liquid-phase enzyme-labelled ligand. CFAbs in the tested sample and solid-phase HLA antigen or HLA antigen of target cell are combined, and simultaneously fixed and existed in enzyme-labelled complement or the enzyme-labelled complement anti body is combined with complement fixed in HLA antigen-CFAb composite, then the correspondent enzyme substrate can be added to produce zymolygical color development reaction.

Description

1. Technical Field [0001] The present invention is based on the methodological basis of immunoenzymatic reaction, and realizes a CDC effect evaluation method for the measurement of CFAbs in the sample by measuring the amount of fixed complement of CFABS. In particular, it relates to the basic measurement principles of HLA serum methodology in organ transplantation immunity. 2. Technical background [0002] Rejection in tissue and organ transplantation is the immune response of the organism's immune system to the foreign transplant (not already). When the protein components (non-antigens) on the cell surface of the transplant come into contact with the recipient's immune system, these non-antigen components are recognized by immunity and stimulate the recipient's humoral immune system and cellular immune system to launch an immune attack on the transplant, destroy and reject it Non-transplanted, resulting in the failure of tissue and organ transplant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/569
Inventor 陈格
Owner PEL FREEZ BIOTECH BEIJING
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