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Monoclonal antibody against toxoplasma gondii as well as preparation method and application thereof

A monoclonal antibody and Toxoplasma gondii technology, applied in biochemical equipment and methods, antibodies, chemical instruments and methods, etc., can solve the problems of difficult acquisition, long time-consuming, high cost, etc., and achieve high accuracy of detection results and good application prospects , good effect of specificity

Active Publication Date: 2015-03-04
SINOVAC BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The second method is not suitable for mass propagation of Toxoplasma gondii
The third method has the disadvantages of high cost, long time consumption, and insufficient number of Toxoplasma gondii in the mass preparation process of Toxoplasma gondii.
Therefore, the second and third methods are not easy to obtain a large number of tachyzoites for the preparation of monoclonal antibodies against Toxoplasma gondii

Method used

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  • Monoclonal antibody against toxoplasma gondii as well as preparation method and application thereof
  • Monoclonal antibody against toxoplasma gondii as well as preparation method and application thereof
  • Monoclonal antibody against toxoplasma gondii as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Immunogen preparation and animal immunization (1)

[0031] 1. Culture Toxoplasma gondii with human embryonic lung fibroblasts (see figure 1 ), and concentrate and purify the harvested Toxoplasma gondii.

[0032] Toxoplasma culture: culture human embryonic lung fibroblasts with MEM containing 8% newborn bovine serum. After the cells grow into a single layer, discard the original medium, add MEM containing 1% newborn bovine serum, inoculate Toxoplasma gondii, and inoculate MOI Toxoplasma gondii was harvested after 7 days. The harvested Toxoplasma gondii was inactivated in a 56°C water bath for 30 minutes.

[0033] Purification of Toxoplasma gondii: let stand for 45 minutes, naturally settle to remove cells and cell clusters; sucrose density gradient centrifugation to remove cell debris: sucrose density gradient centrifugation: first add 700ml of Toxoplasma gondii sample, then add 500ml of low-sugar solution, then add 500ml of high-sugar solution, Wherein the ...

Embodiment 2

[0036] Example 2 Immunogen preparation and animal immunization (2)

[0037] 1. Human embryonic lung fibroblasts were used to culture Toxoplasma gondii, and the harvested Toxoplasma gondii was concentrated and purified.

[0038] Toxoplasma gondii culture: Culture human embryonic lung fibroblasts with MEM containing 9% newborn bovine serum, discard the original medium after the cells grow into a single layer, add MEM containing 2% newborn bovine serum, inoculate Toxoplasma gondii, and inoculate MOI of Harvest Toxoplasma gondii after 0.5, 5 days. The harvested Toxoplasma gondii was inactivated in a 56°C water bath for 30 minutes.

[0039] Purification of Toxoplasma gondii: let stand for 30 minutes, naturally settle to remove cells and cell clusters; remove cell debris by sucrose density gradient centrifugation: sucrose density gradient centrifugation: first add 700ml of Toxoplasma gondii sample, then add 500ml of low-sugar solution, then add 500ml of high-sugar solution, Wherei...

Embodiment 3

[0042] Example 3 Immunogen preparation and animal immunization (3)

[0043] 1. Human embryonic lung fibroblasts were used to culture Toxoplasma gondii, and the harvested Toxoplasma gondii was concentrated and purified.

[0044] Toxoplasma gondii culture: culture human embryonic lung fibroblasts with MEM containing 10% newborn bovine serum, discard the original medium after the cells grow into a monolayer, add MEM containing 2% newborn bovine serum, inoculate Toxoplasma gondii, and inoculate MOI of Harvest Toxoplasma gondii after 0.1, 6 days. The harvested Toxoplasma gondii was inactivated in a 56°C water bath for 30 minutes.

[0045] Purification of Toxoplasma gondii: let stand for 60 minutes, naturally settle to remove cells and cell clusters; remove cell debris by sucrose density gradient centrifugation: sucrose density gradient centrifugation: first add 700ml of Toxoplasma gondii sample, then add 500ml of low-sugar solution, then add 500ml of high-sugar solution, Wherein ...

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Abstract

The invention provides a monoclonal antibody against toxoplasma gondii as well as a preparation method and an application thereof and belongs to the field of biological products. According to the monoclonal antibody, the toxoplasma gondii is cultured by virtue of human embryonic lung fibroblasts, harvested, purified and is repeatedly subjected to freeze thawing, mice are immunized by the freeze-thawed toxoplasma gondii, spleen and myeloma cells of the immunized mice are subjected to SP2 / 0 fusion and screening to obtain a hybridoma cell strain capable of stably secreting the monoclonal antibody against toxoplasma gondii, the hybridoma cell strain has a preservation number of CGMCC No. 9559, the monoclonal antibody secreted by the hybridoma cell strain has high titer and good specificity and can be used for the detection of toxoplasma gondii and can also be used for preparation of kits or diagnostic reagents for detection of toxoplasma gondii, the detection results has high accuracy and the monoclonal antibody has good application prospects in the field of diagnosis of toxoplasma gondii.

Description

technical field [0001] The present invention relates to the fields of biological products and vaccinology, and in particular relates to an anti-toxoplasma monoclonal antibody, a hybridoma cell line and application for producing the antibody, and a method for preparing the monoclonal antibody. Background technique [0002] Toxoplasma gondii, also known as Toxoplasma gondii, is distributed globally and can cause widespread epidemics in humans and various animals, causing serious harm to both humans and animals. The infection rate of Toxoplasma gondii varies from place to place. In many countries and regions, the infection rate is 25% to 50%, and the highest is over 80%. It is estimated that the global average infection rate is about 25%, accounting for a quarter of the world's population. The infection rate of Toxoplasma gondii in our country is 4%-9%. It is listed as the primary pathogen of "TORCH syndrome". [0003] At present, there are many diagnostic and detection method...

Claims

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Application Information

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IPC IPC(8): C07K16/20C12N5/20G01N33/577G01N33/569A61K39/395A61P33/02
Inventor 童钦张星星姚志东刘可心高强尹卫东
Owner SINOVAC BIOTECH
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