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An avidin/ streptavidin magnetic complex particle and the process thereof and purpose thereof

A composite particle and avidin technology, which is applied in the field of preparation of avidin/streptavidin magnetic composite particles, can solve the problems of low biomolecule enrichment ability, biomolecule denaturation and inactivation, low sensitivity, etc., and saves energy. Biological reagent, high biotin-binding activity, effect of improving sensitivity

Active Publication Date: 2007-04-25
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a kind of avidin / streptavidin magnetic composite particle and its preparation method and application, which solves the problem of low labeling amount and relatively low enrichment ability of biomolecules in the background technology, so that the detection The sensitivity of the biomolecules is relatively low, and the direct labeling of biomolecules is likely to cause partial denaturation and inactivation of biomolecules.

Method used

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  • An avidin/ streptavidin magnetic complex particle and the process thereof and purpose thereof
  • An avidin/ streptavidin magnetic complex particle and the process thereof and purpose thereof

Examples

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preparation example Construction

[0038] The avidin / streptavidin biomolecule 2 of the present invention is immobilized on the assembled gold-magnetic composite particle 1 by a simple physical adsorption one-step method. The steps of the preparation method of avidin / streptavidin magnetic composite particles are as follows

[0039] 1. Take 100 μl, 5 mg of assembled gold-magnetic composite particles with a particle size of 0.05-100 μm.

[0040] 2. Equilibrate with balanced salt solution

[0041] 1) Putting the balanced salt solution into a centrifuge tube, so that the assembled gold-magnetic composite particles are dispersed in the balanced salt solution;

[0042] 2) Perform magnetic separation with a magnetic separator, and discard the supernatant.

[0043] Balanced salt solution can use 0.02-0.2M tris-hydrochloric acid buffer Tris-HCl, pH=7.0-7.6; or 0.02-0.2M phosphate buffer PBS, pH=7.0-7.6; or N -2-Hydroxyethylpiperazine-N'-2-ethanesulfonate buffer HEPES, etc. It is better to add 0.1-1.0M NaCl to the bal...

Embodiment 1

[0051] Embodiment 1, the preparation method of streptavidin magnetic composite particle of the present invention

[0052] 1. Take 0.02M Tris-HCl buffer solution, which is added with 0.15M NaCl.

[0053] 2. 100 μl and 5 mg of assembled gold-magnetic composite particles were equilibrated three times with buffer.

[0054] Equilibrium process: Disperse the assembled gold-magnetic composite particles in a balanced salt solution, and discard the supernatant after magnetic separation.

[0055] 3. Add 0.5mg / ml streptavidin biomolecules with high biotin binding activity 300ul, shake and mix well, and react at a constant temperature of 37°C for 20 minutes.

[0056] 4. Discard the supernatant by magnetic separation, and wash with 500 μl 0.02 tris-hydrochloric acid buffer solution, pH=7.4 containing 10 mM EDTA disodium salt and 0.15 M NaCl

Embodiment 2

[0057] Example 2, Streptavidin Magnetic Composite Particles of the present invention are used to link biotinylated oligonucleotide probes

[0058] 1) Take 500 μl of 0.01M Tris-Hcl buffer containing 1M NaCl,

[0059] 2) Equilibrate 500 μg of streptavidin magnetic composite particles. Equilibrate three times, and discard the supernatant after magnetic separation.

[0060] 3) Add 200 μl of biotinylated oligonucleotide probe, that is, 1.5 nmol, use the same amount of probe as a blank to measure the OD value of the solution before binding, and place it at room temperature for 20 minutes;

[0061] 4) Keep the supernatant for magnetic separation to measure the OD value of the combined solution, wash three times with 0.01M Tris-Hcl buffer solution containing 0.15M NaCl, and use magnetic separation to keep the supernatant for measuring the OD value after washing.

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Abstract

An amicable element / link amicable element magnetic compound particle and its Process for preparing and usage are disclosed. The product contains the assembled gold-magnet compound particle in inner layer and the amicable element / link amicable element biomolecule which is coating in outer layer. The assembled gold-magnet compound particle is balanced with balancing brine solution, the high bioepiderm combined with the active amicable element / link amicable element biomolecule is oscillated and mixed at constant temperature, reacted, magnetic separated and cleaned. The invention resolves the technical matters of poor versatility, few marks, deactivation and denaturation of biomolecule in background technique. The invention can be used in marking the biotin biomolecule and the biotin non-biological material directly and then enriching, separating, scavenging and detecting of the biomolecule and non-biomolecule. The product has the property of biological activity, and can improve the sensitivity and specificity of the bioanalysis and detection. The process for preparing is simple, and save a good deal of precious biological agents. The measuring process is easier.

Description

technical field [0001] The invention relates to an avidin / streptavidin magnetic composite particle, a preparation method and application thereof. It specifically relates to an avidin used for labeling any biotinylated molecule such as biotinylated nucleic acid, antigen, antibody, enzyme, polypeptide, polysaccharide, etc., and used for enrichment, separation and detection of various biological and non-biological molecules after labeling Preparation and application of streptavidin magnetic composite particles; or magnetic markers for proteins, nucleic acids, cells, etc., as a rapid enrichment method for the separation, purification and detection of various biological and non-biological molecules Preparation and application of whedin / streptavidin magnetic composite particles. Background technique [0002] The Chinese patent "An assembled magnetic composite particle and its preparation method and application" on August 14, 2003, patent application number 03153486.4, announcemen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/543C12Q1/68
Inventor 崔亚丽陈超张志锋崔婷唐一通张彩峰
Owner XIAN GOLDMAG NANOBIOTECH
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