Nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as preparation method and detecting method of same

A technology of triiodothyroid and nano-magnetic particles, which is applied in the biological field, can solve the problems of high preparation cost and use cost, limit the precision of the detection method, and limit the detection effect, and achieve cost advantages, good accuracy, and improve the detection effect. Effect

Active Publication Date: 2015-07-01
SUZHOU HAOOUBO BIOPHARML
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation cost and use cost of the kit are high, because, on the one hand, when preparing the magnetic particle suspension coated with diiodothyronine-gelatin, not only the process is complicated, but also the two The coating rate of iodothyronine-gelatin coating on magnetic particles is low, resulting in higher cost; on the other hand, it adopts horseradish peroxidase-labeled triiodothyronine antibody, which The preparation is also very cumbersome, and the labeling rate is low, which limits its detection effect and increases the cost
In addition, the uncontrollable and poor stability factors in the preparation process of the kit not only lead to the problem of increased cost as mentioned above, but also make the difference between batches of detection large, which limits the precision of the detection method

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
  • Nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as preparation method and detecting method of same
  • Nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as preparation method and detecting method of same
  • Nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as preparation method and detecting method of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1 The preparation of the first reagent

[0045] (1) Materials and instruments: triiodothyronine (T3) monoclonal antibody (purity over 95wt%, concentration 2mg / mL) preserved in phosphate buffer; fluorescein isothiocyanate (FITC), carbonic acid Reagents such as sodium should be chemically pure; G-25 gel purification column was purchased from GE.

[0046] (2) Preparation steps:

[0047] ① Prepare 0.5 mg / mL FITC solution with 0.1-0.2 mol / L carbonate buffer solution with pH 9.0-10.0;

[0048] ② Add the FITC solution prepared in step ① to the antibody solution according to the ratio of triiodothyronine monoclonal antibody to FITC molecular ratio of 1:20, mix well, and let stand at room temperature for 12 hours to generate T3 antibody-FITC conjugate ;

[0049] ③Separate the reaction liquid after step ② through G-25 gel column, remove unreacted FITC, and obtain a solution containing T3 antibody-FITC conjugate (ie, FITC-labeled T3 antibody);

[0050] ④ Dilute the ...

Embodiment 2

[0051] Embodiment 2 Preparation of the second reagent

[0052] (1) Materials and instruments: T3 antigen (solid powder, purity over 95wt%); alkaline phosphatase preserved in phosphate buffer (ALP solution, ALP purity is about 99%, specific activity is about 1500U / mg, concentration is 10 mg / mL); the cross-linking agent DSS was purchased from THERMO Company, and chemical reagents such as TRIS should be chemically pure; the G-25 gel purification column was a product of GE Company.

[0053] (2) Preparation steps:

[0054] ① Take 1 mg of T3 antigen, add DMSO to dissolve the antigen to a concentration of 20-50 mg / mL, add DSS 0.5 mg, react at room temperature for 2 hours, dilute the reaction solution 1:10 with DMSO, and store it at 2-8 °C for later use;

[0055] ②Take 1mg of ALP solution, with 0.1M NaHCO pH9.5 3 Buffer Dilute the ALP solution to 1mg / ml, add the T3-DMSO solution prepared in step ① to the diluted ALP buffer for the ligation reaction, add the T3-DMSO solution to 1 / 20 ...

Embodiment 3

[0057] The preparation of embodiment 3 magnetic separation reagents

[0058] (1) Materials and instruments:

[0059] Suspension of magnetic particles: the content of magnetic particles is 5wt%, and the magnetic particles contain carboxyl (COOH) active groups. The carboxyl group content per gram (g) of magnetic particles (dry weight) is not less than 0.4 millimoles (mmol), with superparamagnetism, The diameter is between 0.5-2μm.

[0060] Anti-FITC antibody: It can be a polyclonal antibody or a monoclonal antibody, with a purity of more than 90% by weight and a dilution titer of more than 1:1 million;

[0061] 2-Morpholinoethanesulfonic acid (MES), carbodiimide (EDC), TRI S, and other reagents should be of chemical purity.

[0062] (2) Preparation steps:

[0063] ①Take 100mg of magnetic particle suspension, magnetically separate to remove the supernatant, and resuspend in 10mL of 0.05mol / L, pH4.5~5 MES buffer;

[0064] ②Add 2~4mg of anti-FITC antibody and suspend at room te...

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
diameteraaaaaaaaaa
Sensitivityaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention relates to a nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as a preparation method and a detecting method of the nanometer magnetic particle chemiluminescence detection kit. The kit comprises a first reagent, a second reagent and a magnetic separating agent, wherein the first reagent is a solution containing a fluorescein labeled triiodothyronine antibody; the second agent is a solution containing an alkaline phosphatase labeled triiodothyronine antigen; and the magnetic separating agent is a suspension containing magnetic particles coated by a fluorescein antibody. According to the invention, the triiodothyronine can be quantitatively detected with lower cost, higher accuracy and higher precision.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a triiodothyronine (T3) nano-magnetic particle chemiluminescence assay kit combining immunomagnetic particle separation technology and chemiluminescence immunoassay technology and its preparation method and detection method . Background technique [0002] Triiodothyronine (3,5,3'-triiodothyronine, T3), an iodotyrosine with a molecular weight of 651, is an important thyroid hormone like thyroxine (T3) . Both T3 and T3 have two forms of bound state and free state in the circulation, and under normal circumstances, a dynamic balance is maintained between the two forms. [0003] The determination of T3 is very meaningful for diagnosing thyroid dysfunction and monitoring hypothyroidism. For patients with hyperthyroidism, the total T3 and total T3 levels are consistent in most patients. However, in some cases, hyperthyroidism is only caused by increased production of T3 (T3...

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
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/78G01N33/531G01N33/533G01N33/535
Inventor 于大为程晓蕾
Owner SUZHOU HAOOUBO BIOPHARML
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap