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Anti-FPV (feline parvovirus) feline genetic engineering antibody ELISA kit and detection method thereof

A genetically engineered antibody and feline parvovirus technology, which is applied in the field of anti-feline parvovirus feline genetically engineered antibody ELISA kits, can solve problems such as the inapplicability of the detection range, and achieve the effect of high sensitivity

Inactive Publication Date: 2019-11-26
CHANGCHUN SR BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently commercially available Biogo cat triple antibody kit does not mention that it can detect anti-feline parvovirus genetically engineered antibodies, and its detection range cannot be used for feline parvovirus genetically engineered antibodies. The detection method and kit of virus genetic engineering antibody are designed to detect the level of anti-feline parvovirus genetic engineering antibody in animals before and after emergency immunization, evaluate their immune function, analyze the effect of animal immunity, judge whether the immunity is successful, and make the course of the disease controllable The effect, to help the diagnosis and prognosis of the disease

Method used

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  • Anti-FPV (feline parvovirus) feline genetic engineering antibody ELISA kit and detection method thereof
  • Anti-FPV (feline parvovirus) feline genetic engineering antibody ELISA kit and detection method thereof
  • Anti-FPV (feline parvovirus) feline genetic engineering antibody ELISA kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Determination of antigen coating concentration and standard curve initial concentration

[0040] The preparation method of the standard antigen FPV-VLPs protein is as follows: respectively inoculate the suspended Sf9 cells with the recombinant baculovirus according to MOI=0.1, and harvest after culturing for 4 days. Centrifuge the Sf9 cells inoculated with FPV antigen directly at 3000rpm for 30min, discard the supernatant, and wash the cells with an equal volume of 25mM NaHCO 3 Suspend, act on ice for 30 minutes, and centrifuge at 3000 rpm for 30 minutes to obtain the supernatant, which is the harvest liquid containing FPV virus-like particles. The hemagglutination titer is determined using pig blood, and the HA is 1:216. It was used as the standard antigen protein in this experiment.

[0041] The preparation method of the standard antibody is as follows: the serum is separated from the blood of cats infected with FPV, and it is identified as an antibody that...

Embodiment 2

[0043] The determination of the optimal working concentration of the goat anti-cat IgG antibody of embodiment 2 horseradish peroxidase labeling

[0044] According to the operation steps of the above ELISA detection method and the determination of the optimal antigen coating concentration of 2 μg / mL and the initial concentration of the standard curve of 0.25 μg / mL, only the HRP-labeled goat anti-cat IgG antibody was diluted to 1:3000, 1:4000, 1 : 5000, 1: 6000, 1: 7000, 1: 8000 were tested, and the rest of the operations remained unchanged. Repeat the test 3 times, according to the OD of negative and positive serum 450 value, measured OD 450 When the value is close to 1, determine the optimal working concentration of HRP-labeled goat anti-cat IgG antibody. That is, when the HRP-labeled goat anti-cat IgG antibody was diluted 1:6000 times, the OD 450 A value close to 1 is determined to be the optimum working concentration, such as figure 2 shown.

Embodiment 3

[0045] Example 3 Preparation of Anti-Feline Parvovirus Feline Genetic Engineering Antibody ELISA Kit

[0046] According to the optimal optimization conditions determined in the previous examples, add the antigen FPV-VLPs protein to the microtiter plate at a working concentration of 2 μg / mL according to 100 μl / well, and place it at 4°C for 12 hours; wash it three times with 0.05% Tween-20 PBS washing solution , add 2% BSA 100 μL / well to seal for 2 hours; pat dry and place in a sealed bag. Pack the enzyme-labeled antibody reagent, positive control sample and negative control sample, rinsing solution, color developing solution, and stop solution into reagent bottles according to the amount used. Assemble the ELISA plate, reagents, and sealing film into a kit, store at 2-8°C, and use it for later use in experiments.

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Abstract

The invention discloses an anti-FPV (feline parvovirus) feline genetic engineering antibody ELISA kit and a detection method thereof and belongs to the technical field of genetic engineering antibodyanalysis and detection. The kit comprises an ELISA plate coated by an antigen and an enzyme-labeled antibody, wherein the antigen is a FPV-VLPS purified antigen, and the enzyme-labeled antibody is a goat anti-cat IgG antibody labeled by horseradish peroxidase. The detection method comprises the following steps: coating the ELISA plate; drawing a standard curve; adding the goat anti-cat IgG antibody labeled by the horseradish peroxidase, carrying out TMB color developing and stop of a stop solution; measuring an OD450 value of each reaction hole; and performing judgment according to judgment standard and the like. The kit and the detection method have very high sensitivity, specificity and stability in detecting anti-FPV (feline parvovirus) feline genetic engineering antibodies, and providestrong support for study in detecting the level of the anti-FPV (feline parvovirus) feline genetic engineering antibodies in animals before and after emergency immunization.

Description

technical field [0001] The invention relates to the technical field of genetic engineering antibody analysis and detection, in particular to an anti-feline parvovirus feline-derived genetic engineering antibody ELISA kit and a detection method thereof. Background technique [0002] Feline parvovirus (FPV), also known as feline panleukopenia virus, feline infectious enteritis virus or feline distemper virus. Feline parvovirus is a member of the family Parvoviridae and the genus Parvovirus. Feline parvovirus can replicate and multiply in various tissue cells such as spleen, heart, kidney, lung, intestine, testis, bone, adrenal gland and lymph nodes of cats and feline animals. Feline parvovirus mainly infects cats and feline animals (such as tigers, leopards, etc.) ) and Mustelidae (mink, ferret), Procyonidae (long-nosed raccoon, raccoon) animals. It can infect cats of all ages, and mainly causes the disease in kittens under 1 year old. Among them, unvaccinated young cats ag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/535G01N33/543
CPCG01N33/535G01N33/543G01N33/56983G01N33/6854G01N2333/015G01N2333/47
Inventor 石晶李雪殷玉和赵健金宏丽张馨月
Owner CHANGCHUN SR BIOLOGICAL TECH
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