Hybridoma for preparation of scophthalmus maximus immunoglobulin monoclonal antibody

A technology of hybridoma cells and monoclonal antibodies, applied in the field of hybridoma cells, can solve the problems of complicated purification methods and the like

Active Publication Date: 2013-08-28
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

There is only one report on the purification method of turbot immunoglobulin (Wei Jianteng et al., 2008). The purification method is relatively cumbersome, and the purpose of the purification is to prepare polyclonal antibodies (polyclonal anti)

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  • Hybridoma for preparation of scophthalmus maximus immunoglobulin monoclonal antibody
  • Hybridoma for preparation of scophthalmus maximus immunoglobulin monoclonal antibody

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Embodiment Construction

[0015] 1. Separation and purification of serum immunoglobulin from turbot

[0016] Blood was collected from the tail vein of normal turbot, left at room temperature for 1 hour, overnight at 4°C, and the serum was separated by centrifugation (5000 r / min, 30 min, 4°C) the next day, aliquoted and stored in a -70°C refrigerator for later use.

[0017] The salting-out method was used to crudely extract the immunoglobulin from turbot serum. After diluting the turbot serum with an equal volume of 0.01 mol / L (pH=7.4) phosphate buffered saline (PBS), the serum was diluted in 0.01 mol / L (pH=7.4) 5.4) Dialyze in phosphate buffer (PB) at 4°C, collect the precipitate by centrifugation (3000 r / min, 30 min, 4°C), dissolve the precipitate in 0.1 mol / L (pH=5.4) PB, and continue to follow the above method After dialysis, the precipitate was collected by centrifugation again and dissolved in 0.1 mol / L (pH=8.6) PB, then further purified by Sephadex-200 gel column (80 cm × 1.5 cm), and purified wi...

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Abstract

The object of the present invention is to provide a hybridoma for preparation of scophthalmus maximus immunoglobulin monoclonal antibody. The hybridoma can prepare scophthalmus maximus immunoglobulin monoclonal antibody stably and efficiently to establish indirect ELISA method. By the method, multiple pathogen specific immunoglobulin can be detected, and the purpose of early diagnosis for pathogens is realized. The immunization effects of various vaccines can be assessed. Data support is provided to develop a correct immune procedure.

Description

technical field [0001] The invention belongs to the technical field of fish molecular immunology, and in particular relates to a hybridoma cell for preparing turbot immunoglobulin monoclonal antibody. Background technique [0002] Fish is the lowest vertebrate with cellular and humoral immune systems, and immunoglobulin is the most important mediator of the body's immune response. When the fish body is stimulated by foreign substances (such as diseases, vaccines, etc.), the corresponding immunoglobulins will be produced in the body. Accordingly, through the detection of specific immunoglobulins, not only can early diagnosis and early warning of diseases be made, but also real-time detection of antibody responses in animals after vaccination can provide a guarantee for effective immunization procedures. [0003] Turbot (Scophthalmus maximus) is a rare fish species imported from my country. Because of its fast growth and tender meat, it has become the main species of industri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20
Inventor 王蔚芳李青梅甘玲玲高淳仁张改平雷霁霖
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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