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A kind of compound and its preparation method and application

A complex and immune detection method technology, applied in the field of complex and its preparation, can solve problems affecting the detection of target antigens, achieve the effect of reducing errors or errors, reducing interference, and wide application range

Active Publication Date: 2021-11-05
东莞市东阳光诊断产品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the existence of anti-streptavidin antibodies in human serum has seriously affected the detection of target antigens in the human body, and no effective solution has been proposed in the prior art

Method used

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  • A kind of compound and its preparation method and application
  • A kind of compound and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1 prepares biotin-saturated streptavidin (SA-B)

[0043] a) Take 0.6ml SA (10mg / ml, 10mM PB, pH 7.5)

[0044] b) Add 0.1ml D-biotin (1mg / ml, 10mM PB, pH 7.5) to SA, and rotate at 37°C for 1h;

Embodiment 2

[0046] Example 2 Detection of Carcinoembryonic Antigen (CEA)

[0047] 1. Main reagents and materials:

[0048] Streptavidin magnetic beads (Nanjing Pangu Gene), concentration 10mg / ml, 100μl / tube (1mg / tube)

[0049] Conjugated antibody: Fapon CEA-23, Fapon CEA-100

[0050] Coupling reagent: activator Traut's (Thermo), SM (PEG) 4 (Thermo)

[0051] Biotin: NHS-PEG12-Biotin (Thermo)

[0052] Samples to be tested: 16 serum samples from clinics, numbered 1, 2, 3~15, 16

[0053] Substrate: AMPPD (Shenzhen Mercate)

[0054] Buffer 10D: 20mM 7.2PBS+0.1%BSA+0.05%TW-20+0.05%PC300

[0055] Tris buffer: 50mM Tris+0.9%NaCl+0.1%Tween20+0.05%Pc300, pH 7.4

[0056] 2. Steps:

[0057] (1) Biotin antibody preparation (Bio-Ab)

[0058] 1. Antibody buffer exchange

[0059] a. Ultrafiltration tube (0.5mL 50K) + 100μg antibody + PBS buffer (10mM, pH 7.2) to 0.4mL;

[0060] b. 8000g centrifugation;

[0061] c. Repeat step b 2 times to recover.

[0062] 2. Conjugate antibody to biotin

[...

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Abstract

The present invention relates to a compound and its preparation method and application. The present invention provides a compound for reducing the interference of anti-avidin antibodies. The compound includes biotin and avidin, and the avidin saturates Conjugate the biotin. The invention also provides its preparation method and its application in immunoassay. Due to the existence of anti-streptavidin antibodies in part of the human body, in the process of in vitro immunodiagnosis, the antibodies, as endogenous interferents, will affect the detection results, and the complex provided by the invention can bind to the antibodies, thereby avoiding the impact of the antibodies on The detection causes interference, reduces the corresponding error or error, and significantly improves the accuracy of the detection results.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to a compound and its preparation method and application. Background technique [0002] Biotin-avidin system (Biotin-Avidin System, BAS) has been introduced into detection techniques such as enzyme-linked immunoassay, reflection immunoassay and fluorescence immunoassay since the late 1970s, which has improved the stability and specificity of these methods and greatly improved the method sensitivity. For the accurate and rapid detection of trace antigens and antibodies, a new way has been opened up, and it has been widely used in the qualitative and quantitative detection of antigens and antibodies. [0003] Discovered in the early 1960s, biotin is widely present in animals and plants, especially in egg yolk and liver. Biotin is composed of an imidazolone ring and a thiophene ring. The imidazolone ring is mainly bound to avidin, and the carboxyl group at the end of the side cha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/36C07K1/107G01N33/53G01N33/543G01N33/58
CPCC07K14/36G01N33/5306G01N33/54326G01N33/581
Inventor 张瑜陈立勇刘仁源顾志鹏邓晓侠熊灿
Owner 东莞市东阳光诊断产品有限公司