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Preparation method and application of monoclonal antibody against immunoglobulin of yellow catfish

A monoclonal antibody, immunoglobulin technology, applied in the field of fish immunology

Inactive Publication Date: 2016-05-25
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report about the monoclonal antibody of peltatus catfish immunoglobulin

Method used

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  • Preparation method and application of monoclonal antibody against immunoglobulin of yellow catfish
  • Preparation method and application of monoclonal antibody against immunoglobulin of yellow catfish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1. Extraction of Peeled catfish immunoglobulin

[0015] Collection of serum of yellow catfish:

[0016] Blood was collected from the tail vein of healthy yellow catfish, left at room temperature for 1 hour, overnight at 4°C, centrifuged at 4000r / min for 15 minutes the next day, and the supernatant was taken and stored at -80°C for later use.

[0017] Isolation and Purification of Peeled Catfish Immunoglobulin:

[0018] Saturated ammonium sulfate (SAS) salting-out method: After diluting the serum of yellow catfish with an equal volume of normal saline, slowly add saturated ammonium sulfate of pH 7.0 to 30% saturation, let stand overnight at 4°C, centrifuge at 10,000r / min for 30min, and take Continue to add saturated ammonium sulfate to the supernatant until the final saturation is 50%. Treat as above, collect the precipitate, dissolve the precipitate in 0.02mol / LPBS (pH7.4), and then dialyze and desalt with PBS at 4°C to obtain the crude extraction of yellow catfish...

Embodiment 2

[0058] 1. Extraction of Peeled catfish immunoglobulin

[0059] Collection of serum of yellow catfish:

[0060] Blood was collected from the tail vein of healthy yellow catfish, left at room temperature for 1 hour, overnight at 2°C, centrifuged at 4000r / min for 15 minutes the next day, and the supernatant was taken and stored at -80°C for later use.

[0061] Isolation and Purification of Peeled Catfish Immunoglobulin:

[0062] Saturated ammonium sulfate (SAS) salting-out method: After diluting the serum of yellow catfish with an equal volume of normal saline, slowly add saturated ammonium sulfate of pH 6.0 to 40% saturation, let stand overnight at 2°C, centrifuge at 10,000r / min for 30min, and take Continue to add saturated ammonium sulfate to the supernatant until the final saturation is 50%. Treat as above, collect the precipitate, dissolve the precipitate in 0.01mol / LPBS (pH7.0), and then dialyze and desalt with PBS at 4°C to obtain the crude extraction of yellow catfish...

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Abstract

The invention discloses a preparation method and application of a monoclonal antibody against the immunoglobulin of yellow catfish. The monoclonal antibody against the immunoglobulin of yellow catfish is produced by a hybridoma cell strain named as hybridoma F6-F2 and preserved in China Center for Type Culture Collection on September 13th, 2015, with an accession number of CCTCC No. C2015152. The monoclonal antibody is used for detection and evaluation of yellow catfish vaccine immunity effect. According to the invention, tri-antibody sandwich enzyme-linked immunosorbent assay is established, can evaluate the immunization effect of a plurality of vaccines and lays a foundation for development and application of the vaccines.

Description

technical field [0001] The invention belongs to the technical field of fish immunology, and in particular relates to a preparation method and an application of a monoclonal antibody of peliodilis immunoglobulin. Background technique [0002] Yellow catfish (Pelteobagrus fulvidraco) is a common and important small economic fish in my country. It is very popular among consumers because of its fresh and tender meat and few spines. The large-scale cultivation of yellow catfish in my country has a history of more than 10 years, mainly concentrated in the two lakes and the Yangtze River Delta. With the rapid expansion of its breeding scale and the continuous improvement of its breeding density, disease problems have become increasingly prominent, among which bacterial diseases are the main ones. Under the high requirements of protecting the water environment and food safety, immunization prevention has become the best means of disease prevention and treatment, and vaccines are it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/42C12R1/91
Inventor 徐洋沈锦玉姚嘉赟尹文林蔺凌云袁雪梅潘晓艺
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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