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Kit for detecting soluble ST2 protein

A kit and soluble technology, applied in the field of immunomedical detection, can solve problems such as operation errors, and achieve the effects of simple luminescence system, wide linear range and good clinical compliance rate.

Pending Publication Date: 2022-04-19
SHANGHAI UPPER BIO TECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patented magnetic particle chemiluminescence detection reagent (CN108152486A) that has been applied in China is manually operated, and there are certain operational errors

Method used

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  • Kit for detecting soluble ST2 protein
  • Kit for detecting soluble ST2 protein
  • Kit for detecting soluble ST2 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] In this embodiment, the tracer marker is acridinium ester, including:

[0083] Preparation 1: Preparation of primary antibody coated with magnetic beads

[0084] (1) Measure 10mg of magnetic beads (average particle size 3μm, JSR company, solid content 10%), suspend in 1ml 0.1M pH5.5 MES buffer, shake and mix for a short time, use magnet or magnetic stand to absorb for 5-10min Afterwards, discard the supernatant, repeat the above washing step 3 times, add 1ml 0.1M pH5.5 MES buffer, and vortex to mix.

[0085] (2) Add 0.4 mg of ST2 monoclonal antibody (Medix, 100682) at a ratio of magnetic beads:antibody mass ratio of 100:4 to coat in 0.1 M MES buffer, and incubate for 2 hours at room temperature with rotation.

[0086] (3) Add 50 μL of 30 mg / ml 1-(3-dimethylaminopropyl)-3-ethyldiimine hydrochloride (EDC), vortex and incubate at room temperature for 0.5 h.

[0087](4) Add 140 μL 0.15M glycine solution and 70 μL 0.15M ethanolamine solution to the coated antibody, rotate ...

Embodiment 2

[0116] This example provides the screening process of the antibody coating ratio during the preparation of the first antibody.

[0117] Compared with the process of Preparation 1 in Example 1, Example 2 changes the mass ratio of the magnetic beads to the antibody in Step (4) of Preparation 1. In Example 2, in step (4) of Preparation 1, 100:3, 100:4, and 100:5 were respectively used as the mass ratio of magnetic beads to antibodies. Combine the three primary antibodies prepared in Example 2 with the secondary antibodies prepared in Example 1 and 2 to obtain three kits, and then cooperate with a fully automatic chemiluminescence analyzer for detection, and each group is repeated twice. Second, the test results are shown in Table 1.

[0118] Table 1 Screening results of the mass ratio of magnetic beads to antibodies during the preparation of the first antibody

[0119]

[0120] The results in Table 1 show that when screening the mass ratio of magnetic beads to antibodies, th...

Embodiment 3

[0122] This example provides the screening process of the antibody and magnetic bead coupling time (ie, coating time) during the preparation of the first antibody.

[0123] Compared with the process of Preparation 1 in Example 1, Example 3 changed the incubation time of step (4) in Preparation 1. In Example 3, in step (4) of Preparation 1, 1 h, 2 h and 3 h were respectively used as the incubation time. Combine the three primary antibodies prepared in Example 3 with the secondary antibodies prepared in Example 1 and 2 respectively to obtain three kits, and then cooperate with a fully automatic chemiluminescence analyzer for detection, and repeat twice for each group. Second, the test results are shown in Table 2.

[0124] Table 2 Screening results of antibody and magnetic bead coupling time

[0125]

[0126]

[0127] The results in Table 2 show that the coupling time of the antibody and the magnetic beads basically reached equilibrium at 2 hours. Therefore, in the prep...

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Abstract

The invention discloses a kit for detecting soluble ST2 protein. The kit comprises a first antibody coated by magnetic beads and a second antibody marked by a tracing marker, the first antibody and the second antibody are anti-ST2 antibodies and are used for identifying different epitopes respectively, and the specific content is shown in the text of the invention. The kit is high in sensitivity, good in repeatability, wide in linear range, high in interference resistance and high in detection speed, a detection result can be obtained within 10 min, and a sufficient basis can be provided for evaluating risk grading and prognosis of patients with acute heart failure and chronic heart failure and guiding doctors to conduct drug treatment.

Description

technical field [0001] The invention belongs to the field of immunomedicine detection, in particular to a kit for detecting soluble ST2 protein. Background technique [0002] Growth-stimulation expressed gene 2 (ST2) is a member of the interleukin-1 receptor family, among which soluble ST2 (sST2) and transmembrane ST2 (ST2L) are the main research objects of heart failure (HF). ST2 plays a key role in immune and inflammatory responses. The 2013 ACC / AHA / HFSA heart failure guidelines pointed out that soluble ST2 is a marker of myocardial fibrosis, which can predict the probability of admission and death in patients with heart failure, and can also add additional predictive value to natriuretic peptides. ST2 has some unique advantages, such as the test results are not affected by the original cardiovascular disease, atrial fibrillation, age, quality index, etc., and the concentration of NT-proBNP increases significantly with the attenuation of renal function (Bayes-Genise et al...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/532G01N21/76
CPCG01N33/6869G01N33/6887G01N33/6893G01N33/577G01N33/54326G01N33/532G01N21/76G01N2333/7155G01N2800/325
Inventor 郑婉丽张瑞石晓强徐建新李福刚
Owner SHANGHAI UPPER BIO TECH PHARMA
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