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A kind of microsphere reagent and its preparation method and application

A technology of microspheres and reagents, applied in the biological field, can solve the problems of system stability and detection sensitivity to be further improved, and achieve the effects of small restrictions, high production efficiency, and simple operation

Active Publication Date: 2022-03-04
AILEX TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has not been optimized for activation, oxidation and coupling methods, and its system stability and detection sensitivity need to be further improved

Method used

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  • A kind of microsphere reagent and its preparation method and application
  • A kind of microsphere reagent and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] 1) Dissolve the microspheres in additive 1, and record it as mixed solution A;

[0073] 2) Dissolve the β2 microglobulin antibody in auxiliary agent 1, and record it as mixed solution B;

[0074] 3) Dissolve the coupling agent in auxiliary agent 1, and record it as mixed solution C;

[0075] 4) Add the mixture A, B, and C into the container and mix for 36 hours at a speed of 35 rpm. After mixing, the concentration of the microspheres is 0.3%, the concentration of the antibody is 0.3%, and the concentration of the coupling agent is 0.5 mg / L. Dispersion, ultrasonic power 35%, ultrasonic on time 2s, ultrasonic off time 5s, total ultrasonic time 20min;

[0076] 5) Add one-tenth volume of additive 2, continue to mix for 3 hours, rotate at 35 rpm, ultrasonically disperse, ultrasonic power is 35%, ultrasonic on time is 2s, ultrasonic off time is 5s, total ultrasonic time is 20min ;

[0077] 6) Add an equal volume of Auxiliary 3 and mix evenly at a speed of 30 rpm, which is ...

Embodiment 2

[0079] 1) Dissolve the microspheres in additive 1, dilute to a concentration of 0.1%, and record it as mixed solution A;

[0080] 2) Dissolve β2 microglobulin antibody in auxiliary agent 1, dilute to a concentration of 0.1%, and record it as mixed solution B;

[0081] 3) Dissolve the coupling agent in auxiliary agent 1, dilute it to a concentration of 1 mg / mL, and record it as mixed solution C;

[0082] 4) Add the mixed solution A, B, and C into the container and mix for 1 hour at a speed of 20 rpm. For dispersion, the ultrasonic power is 10%, the ultrasonic on time is 3s, the ultrasonic off time is 3s, and the total ultrasonic time is 5min;

[0083] 5) Add one-ninth volume of additive 2, continue mixing for 1 hour, rotate at 20 rpm, ultrasonically disperse, ultrasonic power is 10%, ultrasonic on time is 3s, ultrasonic off time is 3s, total ultrasonic time is 5min ;

[0084] 6) Add ten-ninth volume of auxiliary agent 3, and mix evenly at a speed of 20 rpm, which is the β2 m...

Embodiment 3

[0086] 1) Dissolve the microspheres in additive 1, and record it as mixed solution A;

[0087] 2) Dissolve the β2 microglobulin antibody in auxiliary agent 1, and record it as mixed solution B;

[0088] 3) Dissolve the coupling agent in auxiliary agent 1, and record it as mixed solution C;

[0089] 4) Add the mixed solution A, B, and C into the container and mix for 72 hours at a speed of 50 rpm. After mixing, the concentration of the microspheres is 0.5%, the concentration of the antibody is 0.5%, and the concentration of the coupling agent is 1.0 mg / L. Dispersion, ultrasonic power 50%, ultrasonic on time 1s, ultrasonic off time 6s, total ultrasonic time 30min;

[0090] 5) Add one-eleventh volume of additive 2, continue to mix for 5 hours, rotate at 50 rpm, ultrasonically disperse, ultrasonic power is 50%, ultrasonic on time is 1 s, ultrasonic off time is 6 s, and the total ultrasonic time is 30min;

[0091] 6) Add 10 / 11 volume of auxiliary agent 3, and mix evenly at a spe...

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Abstract

The invention provides a microsphere reagent and its preparation method and application. By optimizing the experimental operation method, creatively mix the microsphere, antibody and coupling agent according to a specific ratio, optimize the concentration and formula of the auxiliary agent, and screen the compounding agent. The requirements of each mixed solution in each step, the type and concentration of stabilizers, storage agents and preservatives are optimized, the auxiliary agents are coordinated with each other, and then ultrasonically mixed, and the synergy of each step and condition is achieved to achieve a uniformly dispersed and stable reagent system. The method is simple and efficient, can effectively reduce production costs, improve production efficiency, and has broad application prospects and huge market value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a microsphere reagent and its preparation method and application. Background technique [0002] β2-microglobulin is a small molecular globulin produced by lymphocytes, platelets, and polymorphonuclear leukocytes, with a molecular weight of 11.8KD. It is the β chain (light chain) part of the human lymphocyte antigen (HLA) on the cell surface (for a single-chain polypeptide), widely exists in plasma, urine, cerebrospinal fluid, saliva and colostrum, the content is very low. The synthesis rate of normal human β2-microglobulin and the amount released from the cell membrane are quite constant, β2-microglobulin can be freely filtered from the glomerulus, and 99.9% is absorbed in the proximal renal tubule, so the determination of blood β2-microglobulin Protein can be used as an indicator to reflect glomerular filtration rate. The detection of β2-microglobulin is considered to be a simple, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531
CPCG01N33/531G01N33/54313
Inventor 李子樵黄磊
Owner AILEX TECH GRP CO LTD
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