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Kit for detecting neurofilament light chain and method and application thereof

A neurofilament light chain and kit technology, which can be applied to measurement devices, analysis by chemical reaction of materials, instruments, etc., can solve the problems of expensive reagent consumables and instruments, difficult to popularize and use NF-L, etc., and achieve detection time. The effect of short, low detection cost and high detection sensitivity

Pending Publication Date: 2022-03-15
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing studies have shown that NF-L can be detected in serum and cerebrospinal fluid, but the content of NF-L in serum is far lower than that in cerebrospinal fluid. Among the clinically applicable products on the market, chemiluminescence kit ELISA kit It can only be used to detect NF-L content in cerebrospinal fluid. If you want to detect NF-L content in serum, you need to use Quanterix’s Simoa series instruments and use the matching single-molecule immunoassay kit. Although this methodology has high sensitivity, However, the reagent consumables and instruments are expensive, which brings difficulties to the popularization and use of NF-L, so there is an urgent need for a highly sensitive method that can detect the content of NF-L in serum

Method used

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  • Kit for detecting neurofilament light chain and method and application thereof
  • Kit for detecting neurofilament light chain and method and application thereof
  • Kit for detecting neurofilament light chain and method and application thereof

Examples

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preparation example Construction

[0081] In one embodiment, a method for preparing the above-mentioned kit is also provided, including:

[0082] Mix the carrier modified with the active group with the coating solution containing the NF-L first antibody, so that the carrier is coated with the NF-L first antibody, and use the blocking solution to react with the carrier coated with the NF-L first antibody , obtaining the NF-L primary antibody immobilized on the carrier;

[0083] The activated small molecule luminescent marker is reacted with the polymer to obtain a signal amplification marker, and the signal amplification marker is activated to react with the NF-L second antibody to obtain a signal amplification marker-labeled NF-L second antibody.

Embodiment 1

[0086] In this example, detection signal amplification technology is used to couple small molecule acridinium esters / isoluminol to macromolecular materials through dehydration condensation, and then the NF-L second antibody is combined with a plurality of acridinium esters / isoluminol Isoluminol is covalently coupled in an indirect manner to amplify the detection signal of NF-L. Among them, the direct luminescence method uses acridinium ester / isoluminol as the signal amplification marker, and the signal amplification marker emits flash under alkaline oxidation conditions, so as to detect the NF-L content according to the signal intensity of the flash.

[0087] In this embodiment, the first antibody of NF-L is: Anti NF-L mAb 47:3 (UD1), the second antibody of NF-L is: AntiNF-L mAb 2:1 (UD2), the first antibody of NF-L , NF-L secondary antibody and antigen were all purchased from Abcam, and the correlation coefficient r≥0.95 was compared with the commercialized NF-lightTM ELISA r...

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Abstract

The invention relates to a kit for detecting a neurofilament light chain and a method and application thereof, the kit comprises a second antibody, the second antibody is used for specifically binding to an antigen to be detected, and the second antibody is modified with a marker for amplifying a signal. The marker on the second antibody is used for amplifying a signal and can directly emit light to generate a detection signal, and the direct light-emitting reaction time is short, so that the NF-L detection speed is greatly improved. In one embodiment, a chemiluminescence method is adopted for immunoassay, the luminescence time is short, the luminescence intensity is in direct proportion to the content of NF-L in the to-be-detected sample, and then the NF-L in the to-be-detected sample is quantified by detecting the luminescence intensity. Therefore, the kit has the advantages of high detection sensitivity, strong specificity, short detection time, low detection cost, automatic operation and the like.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for detecting neurofilament light chains and its method and application. Background technique [0002] Immunomagnetic particle separation technology is an immunological technology based on antigen-antibody specific binding. It mainly depends on the modification groups marked on the surface of magnetic particles, such as amino, carboxyl, tosyl, streptavidin and other groups, which are covalently or non-covalently coupled with labeled antibodies, and can be used to bind specific antigens , in the automatic immunochemiluminescence analyzer, the separation of the corresponding antigenic substances is carried out. [0003] Neurofilament is the main cytoskeletal component of nerve cells. It is important to maintain the integrity of axon caliber and shape, affecting the speed and accuracy of nerve transmission. There are three distinct neurofilament chains, defined by their ...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/543
CPCG01N21/76G01N33/54326
Inventor 陈小茹梁丽雯吴向东
Owner SHENZHEN AMTECH BIOENGINEERING LTD INC
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