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Universal polyclonal antibody of freshwater fish as well as preparation method and application of the universal polyclonal antibody

A polyclonal antibody and freshwater fish technology, which is applied in the fields of biochemistry and molecular immunology, can solve the problems of no commercialization of rabbit anti-fish IgM antibody, and an immunological detection method that hinders the immune response level of freshwater fish.

Inactive Publication Date: 2017-04-19
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, although there have been relevant literature reports on the preparation of antibodies from a single species of fish, there is no report on the preparation of rabbit antibodies by mixing multiple fish antibodies, and there is no commercial commercialization of rabbit anti-fish IgM antibodies on the market. To a certain extent, it hinders the detection of freshwater fish immune response level and the establishment of its immune detection method

Method used

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  • Universal polyclonal antibody of freshwater fish as well as preparation method and application of the universal polyclonal antibody
  • Universal polyclonal antibody of freshwater fish as well as preparation method and application of the universal polyclonal antibody
  • Universal polyclonal antibody of freshwater fish as well as preparation method and application of the universal polyclonal antibody

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation and Effect Detection of Polyclonal Antibody

[0031] 1. Preparation of serum

[0032] Methods: 15 healthy grass carp, crucian carp, carp, bighead carp and tilapia were selected each, blood was collected from the tail vein, left at room temperature for 1 hour, overnight at 4°C, centrifuged at 4000r / min at 4°C for 20 minutes, aliquoted, and stored at -80°C.

[0033] 2. Separation and purification of five fish immunoglobulins

[0034] In this experiment, recombinant protein A (rPeoteinA) affinity chromatography column was used to purify fish immunoglobulin.

[0035]1. Take 40ml of fish serum, dilute it with equal volume of equilibration buffer PBS, centrifuge at 4000r / min at 4°C for 20min, take the supernatant and process it with a 0.45um filter;

[0036] 2. Equilibrate the column with 10 times column volume equilibration buffer PBS and load the sample at a flow rate of 1ml / min;

[0037] 3. In order to fully combine IgM and rProteinA, the sample wa...

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Abstract

The invention belongs to the fields of biological chemistry and molecular immunology and mainly relates to a universal polyclonal antibody of a freshwater fish as well as a preparation method and application of the universal polyclonal antibody. On one hand, the invention provides an immune composition containing grass carp IgM, bighead carp IgM, crucian carp IgM, carp IgM and tilapia IgM. The immune composition provided by the invention is used as an immune antigen for immunizing an animal to obtain the polyclonal antibody with the titer of 1:1000000 or above, so that the polyclonal antibody not only can be applied to detection of immune response levels of the five fishes including a grass carp, a bighead carp, a crucian carp, a carp and a tilapia, but also can be applied to detection of immune response levels of other eight finishes.

Description

technical field [0001] The invention belongs to the fields of biochemistry and molecular immunology, and mainly relates to a general-purpose freshwater fish polyclonal antibody and its preparation and application. Background technique [0002] Immunoglobulin is an important molecule in the humoral immune response of animals, and the main immunoglobulin of teleosts is IgM, which is a tetramer, and each monomer contains 2 heavy chains and 2 light chains. Highly purified fish IgM plays an important role in fish immune research, such as preparing monoclonal antibodies, polyclonal antibodies, detecting fish pathogens and fish immune response levels; exploring the formation and distribution of IgM can also be used for Study the evolution of the immune system. [0003] Freshwater fish are closely related to humans because of their important nutritional value and ornamental value. The pathogens carried by freshwater fish pose a serious threat to human health. At the same time, the ...

Claims

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Application Information

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IPC IPC(8): C07K16/18G01N33/53G01N33/543
CPCG01N33/53G01N33/543G01N33/54313C07K16/18G01N2333/4603
Inventor 余新炳周心怡姜红烨唐泽丽孙恒昌黄艳陈庭金林志鹏任鹏丽
Owner SUN YAT SEN UNIV