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Homogeneous detection kit for interleukin 6 and application thereof

A technology for detecting kits and interleukin, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of short linearity, short shelf life, environmental pollution of radioimmunoassay, etc., and achieve the effect of wide detection range and appropriate sensitivity

Pending Publication Date: 2021-07-16
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing heterogeneous immunoassay methods require cumbersome washing steps; at the same time, it is necessary to overcome the defects of radioimmunoassay environmental pollution, short shelf life, and poor repeatability of enzyme immunoassay
[0004] In the homogeneous immunoassay technique, there is no need for separation and washing steps, which can significantly improve the accuracy of detection and shorten the detection time. However, the currently reported kits for detecting interleukin-6 based on homogeneous chemiluminescence, such as CN106124769A, have the following technical problems: ( 1) The coupling efficiency of the antibody to the photosensitive particle is low; (2) The photosensitive particle coated with the antibody directly reacts with the luminescent particle coated with the antibody as a reagent. Compared with the antibody, the particle is bulky and forms a steric hindrance that makes the reaction rate low. Insufficient short-term response and short linearity; (3) Difficult to balance high sensitivity and wide detection range at the same time

Method used

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  • Homogeneous detection kit for interleukin 6 and application thereof
  • Homogeneous detection kit for interleukin 6 and application thereof
  • Homogeneous detection kit for interleukin 6 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] The preparation of embodiment 1 reagent 1

[0088] 1.1 Preparation and characterization process of the first carrier

[0089] 1) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol methacrylic acid, 10ml water, stir for 10min and pass N 2 30min;

[0090] 2) Weigh 0.12g of potassium persulfate and 0.2g of sodium chloride, dissolve them in 40ml of water to prepare an aqueous solution. This aqueous solution is joined in the reaction system of step 1), continues logical N 2 30min;

[0091] 3) The temperature of the reaction system was raised to 70° C., and the reaction was carried out for 15 hours;

[0092] 4) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water for several times by centrifugal sedimentation until the conductivity of the supernatant at the beginning of centrifugation is close to that of deionized water, then diluted with water, and preserved in ...

Embodiment 2

[0115] The preparation of embodiment 2 reagent 2

[0116] 2.1 Add IL-6 antibody II (monoclonal antibody) to 0.1M NaHCO 3 Buffer, the measured concentration was 1.2mg / ml.

[0117] 2.2 Take 0.5 mg of IL-6 antibody II in step 1), adjust the concentration to 1 mg / ml, add 2.7 μl of biotin with a concentration of 16.8 mg / ml, mix well, and react at 2-8°C for 14-18 hours.

[0118] 2.3 After the reaction is completed, dialyze in 20 mM Tris buffer.

Embodiment 3

[0119] The preparation of embodiment 3 reagent 3

[0120] 3.1 Preparation of the second carrier

[0121] a) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min.

[0122] b) Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride and dissolve them in 40ml of water to prepare an aqueous solution. This aqueous solution is joined in the reaction system of step a), continues to pass through N 2 30min.

[0123] c) The temperature of the reaction system was raised to 70° C. and reacted for 15 hours.

[0124] d) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water for several times by centrifugation and sedimentation until the conductivity of the supernatant at the beginning of centrifugation is close to that of deionized water, then diluted with water, and stored in the form of emulsion.

[0125] e) The ...

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Abstract

The invention relates to a homogeneous detection kit for interleukin 6 and an application thereof. The kit comprises a reagent 1 and a reagent 3, the reagent 1 comprises a first buffer solution and acceptor particles suspended in the first buffer solution, the reagent 3 comprises a second buffer solution and donor particles suspended in the second buffer solution, the acceptor particles comprise a first carrier, the first carrier is filled with a luminous composition, and the particle size distribution variable coefficient C.V value of the acceptor particles in the reagent 1 is not lower than 5% and not higher than 20%; and the donor particles comprise a second carrier, the second carrier is filled with a sensitizing agent, and the sugar content of the donor particles in each milligram of the donor particles in the reagent 3 is not higher than 40 mg. The kit provided by the invention can be used on a chemiluminescence analyzer to monitor the immune state and inflammatory response of an organism, and has relatively proper sensitivity and a very wide detection range.

Description

technical field [0001] The invention belongs to the technical field of homogeneous detection, and in particular relates to a homogeneous detection kit for interleukin-6 and its preparation method and application. Background technique [0002] Interleukin 6 (IL-6) is a pleiotropic cytokine with a wide range of functions, which has physiological activities on B cells, T cells, hematopoietic stem cells, liver cells and brain cells. Interleukin 6 is expressed by a single gene, which contains a sequence of 212 amino acids, and the amino terminal is easily cleaved to form a polypeptide of 184 amino acids, with a molecular weight of about 22-27kD. Clinically, it will rise rapidly during acute inflammatory reactions such as internal and external trauma, surgery, and infection. It also plays a role in chronic inflammation such as rheumatoid arthritis, and can also be used to evaluate the monitoring and prognosis of sepsis. It is mainly used for It is used to monitor the immune statu...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543
CPCG01N33/577G01N33/54313Y02A50/30
Inventor 李坚杨阳李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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