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A preparation method of monoclonal antibody used to replace anti-pika secondary antibody

A monoclonal antibody and anti-pika secondary antibody technology, which is applied in chemical instruments and methods, instruments, peptides, etc., can solve the problems of high cost and poor stability of the second antibody, and achieve strong response, good stability, and high potency high effect

Active Publication Date: 2021-09-03
SHANDONG LVDU BIO SICIENCE & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For this reason, the embodiment of the present invention provides a method for preparing a monoclonal antibody used to replace the anti-pika secondary antibody, so as to solve the problems of poor stability and high cost of the second antibody in the prior art

Method used

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  • A preparation method of monoclonal antibody used to replace anti-pika secondary antibody
  • A preparation method of monoclonal antibody used to replace anti-pika secondary antibody
  • A preparation method of monoclonal antibody used to replace anti-pika secondary antibody

Examples

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Embodiment 1

[0019] Example 1. Preparation of monoclonal antibody used to replace anti-pika secondary antibody

[0020] 1. Preparation of whole antigen immunogen from immune serum

[0021] (1) Wash 1ml of SPA agarose gel prepacked column (Shandong Lvdu Biotechnology Co., Ltd., LD-CH-02) with 5-10 column bed volumes of distilled water;

[0022] (2) Equilibrate 5 to 10 column bed volumes with binding buffer binding buffer (0.02M phosphate buffer, 0.01M NaCl, pH 7.4);

[0023] (3) Dilute 5ml of rabbit serum to 50ml with binding buffer, filter and load with 0.45μm filter membrane;

[0024] (4) Wash 5 to 10 column bed volumes with binding buffer;

[0025] (5) Elute the antibody with elution buffer (0.1M citric acid buffer, pH 4.0), collect the elution peak, and adjust its pH to neutral with neutralization buffer (1M Tris-HCl, pH 9.0), determine Protein concentration, used as an immunogen for backup;

[0026] (6) After each use, regenerate and wash with 3-5 column bed volumes of 0.1M NaOH so...

Embodiment 3

[0043] Example 3, the application of the monoclonal antibody used to replace the anti-pika secondary antibody in the immunochromatographic detection kit

[0044] 1. Application of the monoclonal antibody of the embodiment of the present invention in the qualitative detection card of chloramphenicol

[0045] 1.1 Labeling of chloramphenicol-binding protein

[0046] With 0.1 mol / L of K 2 CO 3Adjust the pH of the colloidal gold to 9.0, add 6 μg of chloramphenicol-binding protein to each 1 mL of colloidal gold solution, shake at 120 rpm for 30 minutes at room temperature, add 20 μL of 10% bovine serum albumin, shake at 120 rpm for 30 minutes at room temperature, centrifuge at 12,000 rpm for 20 minutes, discard the supernatant, The precipitate was dissolved with 0.01MPBS to obtain the labeled chloramphenicol-binding protein.

[0047] 2. Determination of quality control C line and detection line T line

[0048] Select the Sartorius Cn140 membrane, respectively draw the whole anti...

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Abstract

The embodiment of the present invention discloses a preparation method of a monoclonal antibody used to replace the anti-pika secondary antibody. The method includes using purified rabbit serum as the whole antigen, and obtaining hybridoma cells through immune reaction and cell fusion screening. process. The embodiment of the present invention provides a method for preparing a mouse non-specific monoclonal antibody, which uses the whole rabbit serum antigen as an immunogen, through immunogen purification, immunization procedures, cell fusion, hybridoma screening and cloning, antibody preparation and purification Obtained and used to replace the anti-pika secondary antibody used in the prior art. The monoclonal antibody prepared in the embodiment of the present invention has a simple preparation method, can be produced in batches, and has good stability. It can be applied to immunochromatographic detection, has high titer and strong reaction, and is suitable for various modes of immunochromatographic reactions.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of antibody preparation, and specifically relates to a preparation method of a monoclonal antibody used to replace the anti-pika secondary antibody. Background technique [0002] Antibodies from primary sources such as mice or rabbits are called primary antibodies, and secondary antibodies are antibodies that can bind to the primary antibody, that is, antibodies, whose main function is to detect the presence of antibodies and amplify the signal of the primary antibody. The secondary antibody is to use the antigenic property of the protein, which is a macromolecule, to immunize the xenogeneic animal, and the immunoglobulin produced by the immune system of the xenogeneic animal is directed against this antibody. The secondary antibody has played a huge role in experiments such as indirect ELISA and immunochromatography. Immuno-chromatography assay (ICA) is a new type of immunoassay m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/42G01N33/577G01N33/558
CPCC07K16/42G01N33/558G01N33/577
Inventor 武玉香沈志强于金枝崔平
Owner SHANDONG LVDU BIO SICIENCE & TECH
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