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Nanobody, nucleic acid molecule and application of anti-human chorionic gonadotropin beta subunit

A technology of chorionic gonadotropin and nano-antibody, applied in anti-hormone immunoglobulin, anti-animal/human immunoglobulin, application, etc., can solve the problem of lack of treatment methods for Down's syndrome, and achieve stable structure and performance , Small molecular weight, high affinity effect

Active Publication Date: 2021-12-03
成都阿帕克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are currently no effective treatments for Down syndrome

Method used

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  • Nanobody, nucleic acid molecule and application of anti-human chorionic gonadotropin beta subunit
  • Nanobody, nucleic acid molecule and application of anti-human chorionic gonadotropin beta subunit
  • Nanobody, nucleic acid molecule and application of anti-human chorionic gonadotropin beta subunit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Preparation of Nanobodies Against Human Chorionic Gonadotropin β Subunit

[0040] 1 Immunity:

[0041] Take healthy adult alpacas, mix the β subunit of human chorionic gonadotropin with adjuvant, and immunize them by intradermal and subcutaneous injection on the back. The immunization program is shown in Table 1. The third booster immunization After the seventh day, the alpaca peripheral blood was collected for the construction of a phage display library.

[0042] Table 1 Alpaca immunization schedule

[0043]

[0044]

[0045] 2 Alpaca lymphocyte isolation:

[0046] Add 6mL of lymphocyte separation solution to a 15mL centrifuge tube, then add an equal volume of whole blood sample, and centrifuge at 800g for 20min at room temperature;

[0047] Carefully suck the leukocytes suspended in the middle layer into a new centrifuge tube, add 2 times the volume of PBS, 800g, and centrifuge at room temperature for 15min;

[0048] Carefully discard the supernatant, add eryt...

Embodiment 2

[0133] This example provides a conjugate containing an anti-human chorionic gonadotropin beta subunit nanobody, the conjugate comprising the anti-human chorionic gonadotropin beta subunit nanobody provided in Example 1 and Coupling part, coupling part agarose microspheres. The preparation method of this conjugate is as follows:

[0134] The NHS-activated agarose was washed 10 times with 0.1M HCl, equilibrated for 5 min each. with coupling buffer 10mMNa 2 HPO 4 , washed 10 times at pH 7.4, added purified anti-human chorionic gonadotropin beta subunit nanobody 4mg / mL NHS-activated agarose microspheres, and reacted at room temperature for 1 h to make the nanobody covalently coupled to NHS-activated agarose Microspheres. Wash three times with coupling buffer and add 1M Tris buffer pH 8.8 to block unreacted reactive groups. with 10mMNa 2 HPO 4 , washed three times at pH 7.4, and obtained the immunoaffinity adsorption material of human chorionic gonadotropin β subunit-B domai...

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Abstract

The invention discloses a nanobody against human chorionic gonadotropin beta subunit, nucleic acid molecule and application, and relates to the technical field of nanobody. The amino acid sequence of the nanobody against human chorionic gonadotropin disclosed in the present invention is shown in SEQ ID NO.1. The nanobody has the activity of specifically binding to the beta subunit of human chorionic gonadotropin, and can be used It is used to detect human chorionic gonadotropin β subunit, which can replace traditional monoclonal antibody and polyclonal antibody; in addition, the nanobody also has the characteristics of small molecular weight, high affinity, stable structure and performance, and can withstand high temperature and high salt Such harsh conditions increase the shelf life of antibodies.

Description

technical field [0001] The present invention relates to the technical field of nanobodies, in particular to a nanobody against human chorionic gonadotropin beta subunit, nucleic acid molecules and applications. Background technique [0002] Human chorionic gonadotropin (HCG) is a glycoprotein produced by the syncytiotrophoblast of the placental chorion. It consists of alpha and beta units bound together by ionic and hydrophobic bonds. The α subunit is shared by the anterior pituitary hormones, which is similar in structure to the α subunit of FSH, LH and TSH. The structure of the β subunit is different from that of the LH and other β subunits, and can be distinguished in terms of immunological activity. Most of the β subunits in blood exist in the form of whole molecules, and the free β subunits (F-13_HCG) account for about 1-5% of HCG. The free β subunit in blood is produced 6 to 8 days after conception, peaks at 50 to 80 days, and then decreases steadily until it stabili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/26C12N15/13G01N33/76
CPCC07K16/26C07K2317/22C07K2317/569G01N33/76G01N2333/59G01N2800/387
Inventor 陈波罗紫豪
Owner 成都阿帕克生物科技有限公司
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