Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same

A chemiluminescence immunoassay and chemiluminescence technology, which is applied in the field of immunoassay medicine, can solve the problems that the spatially resolved fluorescence method is easily interfered by the environment, needs, and restricts its popularization and use.

Inactive Publication Date: 2009-08-26
北京科美东雅生物技术有限公司
View PDF0 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods all have unavoidable shortcomings: such as quartz crystal microbalance and ICP-MS are expensive, spatially resolved fluorescence method is susceptible to environmental interference, photometric analysis method has low detection sensitivity, and capillary electrophoresis has poor reproducibility. Electrochemical immunoassays require special equipment
It has been reported in the literature that the signal amplification principle of the biotin-avidin system is used to realize the fluorescent immunodetection of AFP, but there is no report on the combination of chemiluminescence detection and magnetic particle system to establish a method with high sensitivity and wide range.
Moreover, the chemiluminescence immunoassay kit in the prior art is a closed fully automatic chemiluminescence measurement system, which requires an expensive fully automatic chemiluminescence measuring instrument, thereby limiting the popularization and use, and cannot be widely used in clinical diagnosis and scientific research work

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same
  • Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same
  • Alpha-fetoprotein magnetic particle chemoluminescence immunoassay kit and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation of the AFP Magnetic Particle Chemiluminescence Immunoassay Assay Kit of the present invention

[0040] 1. Preparation of AFP calibrator

[0041] Dilute the pure AFP product with horse serum into a calibrator, and distribute 0 μg / L, 15 μg / L, 50 μg / L, 150 μg / L, 500 μg / L, 1000 μg / L in 6 bottles.

[0042] 2. Preparation of anti-FITC monoclonal antibody coated magnetic particle solution

[0043] Activate the magnetic particles with a particle size of 2-3 μm with glutaraldehyde, stir at room temperature, mix for 2 hours, apply a magnetic field, let stand for 20-25 minutes, pour out the supernatant, and use 0.01mol / L with a pH value of 7.4 Wash with phosphate buffer three times, and suspend with this solution, the concentration is 50-100 mg / mL; add 60-100 μg of anti-FITC monoclonal antibody to each ml of suspension, stir overnight at 4°C, add a magnetic field, and let it stand for 10 ~ 15min, pour out the supernatant, and block with phosphate buffer (pH...

Embodiment 2

[0070] Embodiment 2 The using method of kit of the present invention

[0071] 1. Sample pretreatment

[0072] The fasting morning serum samples were taken from people, centrifuged at 3000rpm for 5min, and the upper serum was taken for analysis.

[0073] 2. Detection method

[0074] Before using this kit for experiments, it is necessary to take out the magnetic particle solution, the mixture of biotin markers and FITC markers, calibrator / sample to be tested, and enzyme-labeled avidin and place them at room temperature for 15-30 minutes to allow them to balance to Room temperature; after that, adjust the incubator or water bath to 37°C; then, prepare a suitable micro-sampler and corresponding tips and check whether the chemiluminescence instrument is working normally.

[0075] The specific operation steps of using this kit to carry out the experiment according to the method of embodiment 2 are as follows:

[0076]After numbering the round-bottom polystyrene test tubes, add 25...

Embodiment 3

[0077] The methodological examination of the kit of the present invention of embodiment 3

[0078] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art, and the results are as follows:

[0079] 1. Determination of kit precision

[0080] (1) Standard precision experiment

[0081] Three batches of the kits prepared in Example 1 were taken for the precision test, and 10 kits were taken from each batch. Measure the 50 μg / L AFP standard substance 5 times with the kit extracted in Example 1. Calculate the coefficient of variation for the measured concentrations. The measurement results of the three batches of kits in Example 1 are shown in Table 1, and the results show that the coefficient of variation is between 3.8% and 7.3%.

[0082] Table 1 AFP standard repeatability experiment

[0083]

[0084] (2) Sample precision experiment

[0085] Three batches of the kits prepared in Example 1 were taken for the...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The invention relates to the medical field of immunoassay, and in particular provides an AFP magnetic particle chemoluminescence immunoassay kit and a method for preparing the same. The kit comprises: 1) an AFP calibrator; 2) a magnetic particle solution enveloped by a fluorescein isothiocyanate (FITC) resisting monoclone antibody; 3) a mixed solution of a biotin-marked AFP monoclone antibody and an FITC-marked AFP monoclone antibody; 4) a horseradish peroxidase marked streptavidin; 5) a chemoluminescence substrate liquid; and 6) a reaction tube. Furthermore, the method for preparing the kit comprises the following steps: 1) preparing the calibrator by using an AFP pure product; 2) preparing the magnetic particle solution enveloped by the FITC antibody; 3) preparing the mixed solution of the biotin-marked AFP monoclone antibody and the FITC-marked AFP monoclone antibody; 4) marking the streptavidin with the horseradish peroxidase; 5) preparing the chemoluminescence substrate liquid; 6) packaging the calibrator, the magnetic particle solution, the mixed solution, the enzyme marked object, the chemoluminescence substrate liquid and the sub packaging tube; and 7) assembling the components into a finished product. The kit has the advantages of simplicity, fastness, flexibility, stability, and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a magnetic particle chemiluminescence immunoassay kit for wide range and highly sensitive determination of α-fetoprotein (AFP) and a preparation method thereof. The kit of the invention combines biotin-avidin immunomagnification technology, immunomagnetic particle separation technology and chemiluminescence immunoassay technology. Background technique [0002] Cancer has posed a serious threat to human survival and health since its birth. At present, the key to tumor treatment lies in early detection and early treatment. However, at present, hospitals diagnose early-stage tumors mainly by imaging and cytopathological diagnostic techniques. Generally, they are diagnosed only after they have obvious space-occupying lesions and clinical symptoms, but this is not the earliest stage of canceration. With the early diagnosis of tumors entering the protein er...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/68G01N21/76G01N33/577G01N33/574
Inventor 王栩张倩云林金明李振甲应希堂胡国茂
Owner 北京科美东雅生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com