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Human cytomegalovirus (HCMV) anti-UL148 polyclonal antibody and preparation method and application thereof

A human cytomegalovirus, polyclonal antibody technology, applied in the medical field, can solve problems such as inability to export

Inactive Publication Date: 2018-06-29
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The presence of UL130 and UL131 in gH / gL / UL148 co-precipitates may simply reflect the association of UL148 with intermediate gH / gL / UL130 and gH / gL / UL131 complexes formed during the assembly of polymeric gH / gL complexes, but cannot efficiently export from the endoplasmic reticulum

Method used

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  • Human cytomegalovirus (HCMV) anti-UL148 polyclonal antibody and preparation method and application thereof
  • Human cytomegalovirus (HCMV) anti-UL148 polyclonal antibody and preparation method and application thereof
  • Human cytomegalovirus (HCMV) anti-UL148 polyclonal antibody and preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0056] 1. Antibody preparation and ELISA determination of its titer

[0057] 1. Immunization of rabbits

[0058] (1) The rabbit used for immunization is a New Zealand white rabbit with a weight of about 2.0kg (purchased from Changzhou Cavens Experimental Animal Co., Ltd., male, 3 months old, and about 2kg in body weight); Take negative serum.

[0059] (2) Wipe the ear veins of rabbits with 75% ethanol. After the ear veins are fully dilated, insert a syringe into the veins to draw 2ml of negative blood. After standing at room temperature for 120min, the whole blood is centrifuged at 6000g for 10min to collect serum.

[0060] (3) For the first immunization, suck the antigen solution formed by fully emulsifying 1ml of 400μg (the content of UL148 recombinant protein in 1ml of antigen solution is 400μg) purified UL148 recombinant protein solution and 1ml of Freund's complete adjuvant into the syringe. Inject subcutaneously at several points, and inject 200 μl at each point.

[0...

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Abstract

The invention discloses a human cytomegalovirus (HCMV) anti-UL148 polyclonal antibody and a preparation method and application thereof. The HCMV anti-UL148 polyclonal antibody is obtained by taking UL148 recombinant fusion protein as an antigen to perform immunization on a rabbit to obtain serum and then performing purification treatment on the serum. The prepared polyclonal antibody has high purity and strong specificity; the UL148 recombinant protein is used for detecting the specificity of the polyclonal antibody; the Western blot result shows that for the HFF (Human Foreskin Fibroblast) infected with the HCMV, expression of UL148 genes is detected through the Western blot; and the polyclonal antibody can specifically recognize UL148 recombinant protein of which the size is about 54.5kDa. The polyclonal antibody disclosed by the invention can be used for development of HCMV diagnostic kits and HCMV related protein detection kits.

Description

technical field [0001] The invention belongs to the field of medical technology, in particular to a human cytomegalovirus anti-UL148 polyclonal antibody and its preparation method and application. Background technique [0002] CMV is an ancient virus that infects most vertebrates, but each specific CMV infects only one species, and there is little evidence that CMV can cross-infect between species. Human cytomegalovirus, which has a large double-stranded DNA herpesvirus genome, infects most humans for life. Infection with human cytomegalovirus causes mild febrile illness in most people, but severe symptoms in immunocompromised patients or newborns. Almost all infection with human cytomegalovirus spreads widely throughout the body, and different cell types such as epithelial cells, endothelial cells, fibroblasts, and smooth muscle cells produce efficient viral infection. In addition, viruses induce a multitude of immune regulatory pathways to disrupt host innate and adaptiv...

Claims

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Application Information

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IPC IPC(8): C07K16/08C07K16/06C07K1/22G01N33/569
CPCC07K16/065C07K16/088G01N33/56983G01N2333/045
Inventor 陈伟杰陈风华王宇
Owner JINAN UNIVERSITY
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