Anti-human statin alpha monoclonal antibody, preparation method thereof, immunodetection reagent and application

A monoclonal antibody and inhibin technology, applied in the direction of anti-animal/human immunoglobulin, anti-growth factor immunoglobulin, measuring devices, etc., can solve the problems of high price and high cost of differential diagnosis

Pending Publication Date: 2022-04-05
河南赛诺特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Monoclonal antibody R1 is an imported reagent, which is expensive, which also leads to high cost of differential diagnosis in this field

Method used

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  • Anti-human statin alpha monoclonal antibody, preparation method thereof, immunodetection reagent and application
  • Anti-human statin alpha monoclonal antibody, preparation method thereof, immunodetection reagent and application
  • Anti-human statin alpha monoclonal antibody, preparation method thereof, immunodetection reagent and application

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Experimental program
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Effect test

Embodiment 1

[0032] The anti-human inhibin α monoclonal antibody of this embodiment comprises a heavy chain variable region with an amino acid sequence shown in SEQ ID NO:7 and a light chain variable region with an amino acid sequence shown in SEQ ID NO:8. The heavy chain variable region comprises VHCDR1, VHCDR2 and VHCDR3 as shown in SEQ ID NO: 1-3, and the light chain variable region comprises VLCDR1, VLCDR2 and VLCDR3 as shown in SEQ ID NO: 4-6.

[0033] Two. Specific examples of the preparation method of the monoclonal antibody against human inhibin alpha

Embodiment 2

[0035] The preparation method of the anti-human inhibin alpha monoclonal antibody of the present embodiment comprises the following steps:

[0036] (1) Rabbit immunization: add 5 kinds of immunogens (immunogens I, II, III, IV, and V are MSH6, Inhibin-α, MLH1, CK19, and PMS2) and immune partners (CSF2 protein; enhance immune effect) into water-soluble In the adjuvant, the immune reagent was obtained, and the concentration of the immune partner in the immune reagent was controlled to be 50 μg / mL, and the immune dose of the five immunogens was 100 μg / monkey. Then, New Zealand white rabbits were immunized with immune reagents, and after the immune effect was evaluated to meet expectations, the rabbits were subjected to immunogen shock immunization, so as to mobilize the rapid activation of B lymphocytes targeting the target immunogen in the rabbit body.

[0037] CSF2 protein can be prepared by a method comprising the following steps:

[0038] 1) First, by consulting the Uniprot d...

Embodiment 3

[0058] The immunological detection reagent of this embodiment is an immunohistochemical detection reagent, which is the monoclonal antibody working solution of Example 1, the concentration of the monoclonal antibody is 0.1 μg / mL, and the solvent is TBS buffer solution, which contains a concentration of 10 mg / mL BSA (bovine serum albumin) and preservatives Proclin-300 and Proclin-950 were added in a ratio of 1:1000 (V / V).

[0059] 4. Specific examples of the application of monoclonal antibodies in immunohistochemistry

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Abstract

The invention belongs to the field of monoclonal antibodies, and particularly relates to an anti-human statin alpha (Inhistatin-alpha) monoclonal antibody as well as a preparation method, an immunodetection reagent and application thereof. The monoclonal antibody comprises a VHCDR1, a VHCDR2 and a VHCDR3 of which the amino acid sequences are shown as SEQ ID NO: 1-3, and a VLCDR1, a VLCDR2 and a VLCDR3 of which the amino acid sequences are shown as SEQ ID NO: 4-6. The anti-human statin alpha monoclonal antibody provided by the invention can accurately identify Inhibin-alpha protein expressed in paraffin tissue and frozen tissue, and compared with an imported reagent R1, the monoclonal antibody has the advantages of consistent specific evaluation result, low antibody use concentration and stronger dyeing effect, and can be used for immunodetection analysis. The monoclonal antibody has important significance for realizing localization of the monoclonal antibody and reducing reagent cost.

Description

technical field [0001] The invention belongs to the field of monoclonal antibodies, and in particular relates to an anti-human inhibin-α (Inhibin-α) monoclonal antibody, a preparation method thereof, an immunodetection reagent and applications thereof. Background technique [0002] Inhibin (INH) is an important regulator of activin (ACT) function. The structure of inhibin is related to activin, and both belong to the transforming growth factor β (TGF-β) superfamily member. Inhibin is a heterodimeric glycoprotein INHA or INHB formed by two subunits of α and β (βA or βB) through disulfide bonds, with a relative molecular weight of 32KD. The respective bases of these three subunits α, βA, and βB The base sequences were located on chromosome 2 (α subunit and βB subunit) and chromosome 7 (βA subunit). Inhibin acts against activin by competing for binding to activin type II receptors, a process that requires inhibin co-receptor-β-glycans to increase inhibin affinity. [0003] In...

Claims

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Application Information

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IPC IPC(8): C07K16/22C12N15/85G01N33/577G01N33/574
Inventor 翟晋豫王远利许鹏程王晓亚李艳娟吴夏梦常秋霜秦志洁齐华李三华
Owner 河南赛诺特生物技术有限公司
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