Method for rapidly typing mammal sperms by using biologically modified beta-cyclodextrin nano magnetic particles

A nano-magnetic particle, magnetic nano-particle technology, applied in the fields of nanotechnology, animal cells, nanotechnology, etc. for materials and surface science, can solve the problems of strong interaction, poor dispersion, low separation efficiency, etc., and achieve a wide range of applications Value, uniform and stable coating effect

Active Publication Date: 2022-07-12
北京田园奥瑞生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can overcome the disadvantages of low separation efficiency, strong interaction between particles and poor dispersion of conventional micron-sized immunomagnetic particles

Method used

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  • Method for rapidly typing mammal sperms by using biologically modified beta-cyclodextrin nano magnetic particles
  • Method for rapidly typing mammal sperms by using biologically modified beta-cyclodextrin nano magnetic particles
  • Method for rapidly typing mammal sperms by using biologically modified beta-cyclodextrin nano magnetic particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation method of nano-magnetic particles

[0031] (1) Preparation of biologically modified β-cyclodextrin nanomagnetic particles:

[0032]Amino-modified magnetic nanoparticles: accurate weighing of FeCl 2 ·4H 2 O (1.25g), add 7.75mL of double distilled water to dissolve, add 6.25mL of ammonia water under vigorous stirring, continue to stir for 10 minutes, then add 2.5mL of 3-aminopropyltriethoxysilane (APTES), at 134 ℃ Autoclave for 3 h at temperature and collect the black magnetic solution. The reaction product was centrifuged at 1000 rpm for 1 minute, after removing large particles, the supernatant was collected, centrifuged at 6000 rpm for 5 minutes, the black precipitate was collected, washed with double distilled water for 3 times, and washed with absolute ethanol twice (to remove the unreacted APTES in the solution). ), the precipitate was washed twice with double distilled water, and finally the magnetic particles were dispersed in 5 mL of doubl...

Embodiment 2

[0037] Example 2 Sperm typing and separation using biomodified β-cyclodextrin nanomagnetic particles

[0038] Select the qualified fresh swine semen as the experimental sample and use the density meter to detect the actual concentration of the original semen, add 100 million fresh semen to 5 ml of 0.1% PBS, mix gently, and centrifuge at 1500 rpm for 7 min at room temperature;

[0039] Remove the supernatant, add 300 μL of 0.01mol / L PBS, 0.5% BSA, pH7.2 adsorption solution, shake and mix well, add an appropriate amount of the TLR7 / TLR8 ligand-coated nanomagnetic particle suspension prepared in Example 1 (3- 5 μl), after mixing, incubate at room temperature for 30 min in the dark;

[0040] After the incubation, place the above centrifuge tube on the magnetic separation rack for 5 minutes to absorb and discard the liquid (reserve part for subsequent experiments); add 300 μL of 0.01mol / L PBS, 0.5% BSA, 2mM EDTA, pH7.2 for washing Liquid washing, magnetic separation rack adsorptio...

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Abstract

The invention provides a method for rapidly typing mammal sperms by using biologically modified beta-cyclodextrin nano magnetic particles, by which a large amount of sperms can be separated even in a short time, and the motility of the separated sperms can be prevented from being reduced. On one hand, the nano magnetic particle can be used as a carrier of biomacromolecules, and because the nano magnetic particle is coated with a TLR7 / TLR8 ligand, the nano magnetic particle not only has the characteristic of specifically recognizing X / Y sperms, but also has the characteristic of magnetism, so that the nano magnetic particle has more advantages in the aspects of separating, purifying and concentrating target sperms from a complex sample and the like; the method has the advantages of rapidness (5-20 minutes), high specificity, simplicity and convenience in operation, wide application range (almost applicable to treatment of all samples) and the like. The defects of low separation efficiency, relatively strong interaction among particles and relatively poor dispersity of conventional micron-sized immunomagnetic particles can be overcome.

Description

technical field [0001] The invention relates to the field of rapid biological separation, in particular to a rapid separation technology of nano-magnetic particles, in particular to a method for rapid separation of mammalian X and Y spermatozoa by cationic polyelectrolyte-coated magnetic nano-composite particles and its application. Background technique [0002] Techniques for sex pre-selection in livestock greatly contribute to the efficient production of livestock. In the pig industry, for example, male barrows grow faster than sows and reach transport weight in a shorter finishing period. An increase in the number of male offspring in pig production will not only improve the management of the pig industry, but also increase pork production for feed, leading to an increase in food production worldwide. However, criticism of castration has grown, and only a small percentage of cattle farmers currently employ artificial insemination methods involving sex-specific sperm cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J13/10B01J13/02B82Y30/00C12N5/076
CPCB01J13/10B01J13/025B82Y30/00C12N5/061C12N2509/00
Inventor 邓晓彬陆建黄凡
Owner 北京田园奥瑞生物科技有限公司
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