Preparation method of ovomucoid rabbit polyclonal antibody and immunoassay method

An ovomucoid and polyclonal antibody technology, applied in the field of macromolecular protein immunization, can solve the problem of less research on polyclonal antibodies, and achieve the effects of high sensitivity and specificity, high titer and good sensitivity

Inactive Publication Date: 2015-09-30
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing ovomucoid antibodies are monoclonal antibodies, and there are few studies on polyclonal antibodies. It is necessary to develop a polyclonal antibody to improve the sensitivity and specificity of the antibody

Method used

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  • Preparation method of ovomucoid rabbit polyclonal antibody and immunoassay method
  • Preparation method of ovomucoid rabbit polyclonal antibody and immunoassay method
  • Preparation method of ovomucoid rabbit polyclonal antibody and immunoassay method

Examples

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Embodiment 1

[0046] A method for preparing a rabbit polyclonal antibody against ovomucoid, characterized in that it includes two steps: an immunization process and an antibody purification process;

[0047] In the immunization process, rabbits are selected for immunization, and the immunogen is ovomucoid. The immunization method adopts subcutaneous and intramuscular injection. After the initial immunization, four booster immunizations are carried out, and the booster immunizations are performed 2 weeks and 4 weeks after the initial immunization And 6 weeks and 8 weeks, one week after the five immunizations, whole blood was collected from the femoral artery and stored.

[0048] The antibody purification process specifically includes the following steps:

[0049] 1) Sonicate all reagents required in the experiment for 20 minutes to remove air bubbles;

[0050] 2) Equilibrate the purification column: first, flush the pipeline for 1-2 hours, the selected reagent is phosphate buffer solution w...

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Abstract

The invention provides a preparation method of an ovomucoid rabbit polyclonal antibody and an immunoassay method. The preparation method comprises the following main steps: directly immunizing a new Zealand white rabbit by using ovomucoid for five times, and then taking whole blood from arteria femoralis, so that an ovomucoid antibody serum is obtained; and through optimizing the coating amount of the antibody, detecting the dilution ratio of the antibody and the pH values of confining liquid and a sample buffer solution, so that an ovomucoid indirect competitive ELISA method is established. The antibody prepared according to the invention is relatively high in potency and relatively good in inhibition rate, and the established method is relatively good in sensitivity and precision.

Description

technical field [0001] The invention belongs to the technical field of macromolecular protein immunity, and in particular relates to a preparation method and an immunological analysis method of ovomucoid rabbit polyclonal antibody. Background technique [0002] Ovomucoid is composed of 186 amino acids, and its molecular mass is about 28DK. Ovomucoid contains three independent homologous domains. These three functional domains are arranged one after the other and are intramolecular The disulfide bonds connect the various functional domains. The ovomucoid molecule contains 9 intramolecular disulfide bonds, in addition to 20% to 25% glycosyl components, and the most allergenic of the three regions is the third region. The content of ovomucoid in egg white is the highest, and the allergenicity is also the strongest. [0003] Ovomucoid has the highest stability due to the glycosyl component contained in ovomucoid. It is quite stable to trypsin degradation, and it is also quite ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06G01N33/68
Inventor 张燕生威王薇刘冰陆旸王硕
Owner TIANJIN UNIV OF SCI & TECH
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