Single sperm freezing carrier in combination with monoclonal antibodies and freezing and unfreezing method

A monoclonal antibody and freezing carrier technology, which can be used in measurement devices, instruments, scientific instruments, etc., and can solve problems such as difficulty in finding sperm.

Inactive Publication Date: 2018-07-17
周高见 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the problems existing in the prior art, the present invention provides a monoclonal antibody combined with a single sperm freezing carrier and a freezing and thawing method, the purpose of which is to fix single or several sperms on the surface of the carrier, thereby preventing the sperm from

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The monoclonal antibody in this example binds to the single sperm cryocarrier, the monoclonal antibody is ACRV1acrosomal vesicle protein 1 [Homo sapiens (human)], the manufacturer of the monoclonal antibody is sant cruz, the article number is sc5163.

[0032] The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.

[0033] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:

[0034] (1) Add the slides one by one into the piranha solution and soak for 15 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 105°C for 30 minutes. spare;

[0035] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 2.5%, soaking the above-mentioned oven-dried slides in t...

Embodiment 2

[0043] The monoclonal antibody of this example is combined with a single sperm freezing carrier, the monoclonal antibody is PH-20 [Homosapiens], the manufacturer of the monoclonal antibody is sant cruz, and the article number is sc5163. The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.

[0044] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:

[0045] (1) Add the slides one by one into the piranha solution and soak for 150 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 80°C for 30 minutes. spare;

[0046] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 10%, soaking the above-mentioned oven-dried slides in the liquid for 72 hours;

[004...

Embodiment 3

[0055] The monoclonal antibody in this example binds to the single sperm cryocarrier, the monoclonal antibody is ACRV1acrosomal vesicle protein 1 [Homo sapiens (human)], the manufacturer of the monoclonal antibody is sant cruz, the article number is sc5163. The volume ratio of monoclonal antibody to sperm is (10-1000):1 to form antibody sperm solution.

[0056] The freezing and thawing method of the monoclonal antibody combined with the single sperm freezing carrier carried out according to the following steps:

[0057] (1) Add the slides one by one into the piranha solution and soak for 240 minutes, then put them into pure water for ultrasonication, change the water at least three times to remove the organic matter on the surface of the slides, and dry the slides in a drying oven at 150°C for 30 minutes. spare;

[0058] (2) Dissolving 3-aminopropyltriethoxysilane (APTES) in absolute ethanol with a mass ratio of 1%, soaking the above-mentioned oven-dried glass slides in the l...

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PUM

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Abstract

The invention relates to the technical field of sperm freezing, in particular to a single sperm freezing carrier in combination with monoclonal antibodies and a freezing and unfreezing method. Firstly, an APTES is utilized to make the outer surface of a slide become a carbanyl group, the carbanyl group very easily reacts with an amino group on the surface of immunoglobulin, then some monoclonal antibodies with a certain characteristic combine with the surface specificity of the sperm, and the adhesive force of the antibodies fixes the sperm to the surface of the slide. The invention provides amethod capable of fixing one or more sperms to the surface of the carrier, and the sperms can be prevented from being lost in the storage, transportation and operation process.

Description

technical field [0001] The invention relates to the technical field of sperm freezing, in particular to a monoclonal antibody combined with a single sperm freezing carrier and a freezing and thawing method. Background technique [0002] In clinical male infertility patients, some patients with spermatogenesis dysfunction can only obtain a very small amount of sperm during the sperm retrieval process, and only through multiple sperm retrieval and frozen sperm can meet the patient's fertility requirements. The damage of freezing will cause the loss of some sperm, and the existing sperm freezing technology is mainly aimed at freezing a large number of sperm, which will cause the loss of some sperm. For patients with spermatogenic dysfunction who need to freeze sperm, the loss of sperm can be fatal. Therefore, there is a need to develop a freezing technique that does not lose sperm. Contents of the invention [0003] Aiming at the problems existing in the prior art, the pres...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/42G01N1/44G01N1/34G01N33/577G01N33/531
CPCG01N1/30G01N1/34G01N1/42G01N1/44G01N33/531G01N33/577
Inventor 周高见许东汤乐朱文兵邢柳
Owner 周高见
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