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Carrier-hapten compound as well as preparation method and application thereof

A technology of haptens and complexes, applied in the field of immunology, can solve the problems of inability to arrange haptens and control the arrangement of haptens, and achieve the effect of improving the immune effect

Inactive Publication Date: 2016-10-12
NORTHEAST FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these protein carriers cannot control the arrangement of haptens on their surface, and cannot arrange the haptens in an orderly manner on their surface, so they cannot effectively cross-link B cell receptors to promote B cell maturation

Method used

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  • Carrier-hapten compound as well as preparation method and application thereof
  • Carrier-hapten compound as well as preparation method and application thereof
  • Carrier-hapten compound as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Extract TMV-EPMK and CPMV (purchased from: Changchun Institute of Applied Chemistry) from infected tobacco leaves, the specific method is as follows:

[0038] (1) Collect fresh or frozen tobacco leaves infected by TMV-EPMK and CPMV at -80°C;

[0039] (2) Homogenize the tobacco leaves infected by TMV-EPMK and CPMV with 0.2% β-mercaptoethanol and potassium phosphate buffer (TMV-EPMK is 0.1M, pH is 7.8, CPMV is 0.1M, pH is 7.0 );

[0040] (3) with 0.1M, pH is 7.0 potassium phosphate buffer solution and 0.2% β-mercaptoethanol to carry out homogenization treatment to the tobacco mosaic leaf of CPMV infection;

[0041] (4) Filter the homogenate with two layers of gauze, centrifuge at 9000rpm at room temperature for 15min, and take the supernatant;

[0042] (5) The supernatant is mixed with chloroform (CHCl3) in a ratio of 1:1;

[0043] (6) Collect TMV-EPMK and CPMV by precipitation on ice with 8% polyethylene glycol and 0.2M sodium chloride solution;

[0044] (7) The col...

Embodiment 3

[0065] Embodiment 3TMV-EPMK-μE 3 , CPMV-μE 3 , KLH-μE 3 and BSA-μE 3 Serum titer comparison of immunized mice

[0066] 1.2.0 μg / mL μE 3 to TMV-EPMK-μE 3 , CPMV-μE 3 , KLH-μE 3 and BSA-μE 3 Comparative detection of antibody titer in the tail blood of immunized mice

[0067] By directly coating μE 3 to TMV-EPMK-μE 3 , CPMV-μE 3 , KLH-μE 3 and BSA-μE 3 The tail blood of the immunized mice was tested for antibody titer comparison, 2.0 μg / mL was used for coating, and the mouse serum was diluted 4× for sample loading. See the results in Figure 10 .

[0068] It can be seen from the results of serum antibody titer that TMV-EPMK-μE 3 The serum titer obtained from the immunized mice was the highest, reaching 10 -3 The maximum OD value reached 1.8, which was significantly higher than the other three carriers, indicating that the rod-shaped TMV was used as a carrier to make μE 3 With stronger immunogenicity, it can stimulate the body's immune system to produce B cells th...

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PUM

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Abstract

The invention provides a carrier-hapten compound and a preparation method thereof. Carriers are lysine mutant tobacco mosaic virus and cowpea mosaic virus. A hapten is estriol. The preparation method of the carrier-hapten compound comprises the steps of (1) extracting plant viruses; and (2) linking the plant viruses TMV-EPMK and CPMV with estriol. The invention further provides application of the carrier-hapten compound in pregnancy examinations of estriol level of pregnant woman and the like.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to a carrier hapten complex and its preparation method and application. Background technique [0002] Hapten refers to a class of substances whose molecular weight is too small to stimulate and cause an effective immune response in the body, but can combine with the corresponding antibody to generate an antigen-antibody reaction. It only has reactogenicity, not immunogenicity. Known as incomplete antigens, haptens are antigens that cannot alone stimulate the human or animal body to produce antibodies, such as polysaccharides, polypeptides, steroid hormones, fatty amines, lipids, nucleic acids, certain drugs (including antibiotics) and other chemical substances are all haptens. Once the hapten is combined with other larger "non-antigenic" carriers, it can stimulate and cause an effective immune response in the body. If the hapten is chemically combined with a pure protein mole...

Claims

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Application Information

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IPC IPC(8): G01N33/74
CPCG01N33/743
Inventor 魏德强赵霞王倩
Owner NORTHEAST FORESTRY UNIVERSITY