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Matrix protein monoclonal antibody for appraising mineralized stimulating factor and appraising method

A monoclonal antibody, matrix protein technology

Inactive Publication Date: 2010-05-19
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of corresponding research models and methods, the progress of studying the mechanism of biomineralization from the perspective of cell and protein regulation is slow, taking pearl shellfish as the object.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The mantle cells of Hepu pinnacle oyster were taken and cultured for 2 days in the normal culture medium by the tissue block method. Then, according to the types of stimulatory factors to be identified, different stimulatory factors were added to the culture medium and cultured for another 2 days. , to obtain the medium containing the matrix protein secreted by the mantle; BALB / c mice were taken for 4 times of immunization, the splenocytes of the immunized mice were fused with myeloma SP2 / 0 cells, and the enzyme-linked immunosorbent assay (ELISA) was used to detect Methods The positive hybridoma cells were screened out, the positive hybridoma cells were cloned, and the monoclonal hybridoma cells that specifically recognized the matrix protein were screened out, and the monoclonal hybridoma cells were injected into normal BALB / c mice to obtain The ascites of the mouse was affinity-purified to obtain monoclonal antibodies; New Zealand rabbits were taken and immunized with ...

Embodiment 2

[0041] The mantle cells of Hepu pinnacle oyster were taken and cultured for 4 days in the normal culture medium by the tissue block method. Then, according to the types of stimulatory factors to be identified, different stimulatory factors were added to the medium and cultured for another 4 days. , to obtain the medium containing the matrix protein secreted by the mantle; BALB / c mice were taken for 4 times of immunization, the splenocytes of the immunized mice were fused with myeloma SP2 / 0 cells, and the enzyme-linked immunosorbent assay (ELISA) was used to detect Methods The positive hybridoma cells were screened out, the positive hybridoma cells were cloned, and the monoclonal hybridoma cells that specifically recognized the matrix protein were screened out, and the monoclonal hybridoma cells were injected into normal BALB / c mice to obtain The ascites of the mouse was affinity-purified to obtain monoclonal antibodies; New Zealand rabbits were taken and immunized with matrix p...

Embodiment 3

[0043] The mantle cells of Hepu pinnacle oyster were taken and cultured for 3 days in a normal culture medium by the tissue block method. Then, according to the types of stimulatory factors to be identified, different stimulatory factors were added to the medium and cultured for another 3 days. , to obtain the medium containing the matrix protein secreted by the mantle; BALB / c mice were taken for 4 times of immunization, the splenocytes of the immunized mice were fused with myeloma SP2 / 0 cells, and the enzyme-linked immunosorbent assay (ELISA) was used to detect Methods The positive hybridoma cells were screened out, the positive hybridoma cells were cloned, and the monoclonal hybridoma cells that specifically recognized the matrix protein were screened out, and the monoclonal hybridoma cells were injected into normal BALB / c mice to obtain The ascites of the mouse was affinity-purified to obtain monoclonal antibodies; New Zealand rabbits were taken and immunized with matrix pro...

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PUM

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Abstract

The invention relates to a matrix protein monoclonal antibody for appraising a mineralized stimulating factor and an appraising method. The monoclonal antibody specifically distinguishes the matrix protein of Pinctada martensi. The appraising method comprises the following steps of: culturing pallium cells of the Pinctada martensi to obtain a culture medium of primary culture; respectively adding stimulating factors required to be appraised in the culture medium, and then culturing to obtain a cell culture medium; adopting a hybridoma method to obtain a mice monoclonal antibody; using the matrix protein to immunize a New Zealand rabbit to obtain a matrix protein polyclonal antibody; using the monoclonal antibody to establish a method for testing high specificity and sensitivity; meanwhile, under the existence of different stimulating factors, testing the secretion change of the matrix protein of the pallium cells of the Pinctada martensi of primary culture. The invention provides a new method for researching the regulation and control of mineralized relevant matrix protein, thereby laying the foundation for the research on mineralized models.

Description

technical field [0001] The present invention relates to an antibody for identifying the mineralization stimulating factor of Hepu pinnacle oyster, in particular to a matrix protein monoclonal antibody for identifying the mineralizing stimulating factor, and the present invention also relates to using the monoclonal antibody to detect the mineralizing stimulating factor method of identification. Background technique [0002] Biomineralization is a process in which mineral ions are transformed into mineral deposits through the life activities of organisms that are ubiquitous in nature. Most of the biominerals play the role of supporting, defending and protecting organisms, and have functions such as photoreceptors, gravity perception, magnetic navigation and ion storage, which are of vital significance to the survival of organisms. Studies have shown that the biomineralization process of shellfish is regulated by the matrix protein (nacrein), and the matrix protein is closely...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/577
Inventor 张荣庆谢莉萍李琪上官俊龙刘晓军周玉娟
Owner TSINGHUA UNIV
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