A human monoclonal antibody against the vp1 protein of jc virus

A monoclonal antibody, JC virus technology, applied in the direction of antiviral immunoglobulins, antibodies, antiviral agents, etc., can solve the problem of not raising or mentioning neutralizing antibodies in any way

Active Publication Date: 2015-04-15
POMONA RICERCA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For this risk stratification, the authors propose three risk factors, namely: presence or absence of anti-JCV antibodies, prior use of immunosuppressants, and duration of treatment with natalizumab, but they do not address or refer to the assessment of the presence of neutralizing antibodies in the human fluid response

Method used

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  • A human monoclonal antibody against the vp1 protein of jc virus
  • A human monoclonal antibody against the vp1 protein of jc virus
  • A human monoclonal antibody against the vp1 protein of jc virus

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Embodiment 1

[0033] By methods similar to those previously described (Plaisant, P., et al., Human monoclonal recombinant Fabs specific for HCV antigens obtained by repertoire cloning in phage display combinatorial vectors. Res Virol, 1997.148(2): p.165-9 ) was constructed in pPD phagemid vector of IgG1 / k isotype and had 2x10 7 A combinatorial phage-display library of human antibody fragments (monovalent Fabs) at an estimated scale of three elements. The library was generated from the bone marrow of a 68-year-old male whose serum tested positive for the presence of anti-VP1 / JCV antibodies by ELISA. By using 100ng / well of recombinant JCV VP1 protein (Mad1, ) to coat a 96-well plate for ELISA. Several serum dilutions were added to VP1 coated plates in duplicate. The plate was incubated at 37°C for 1 hour and then washed with 0.1% PBS / TWEEN 20. Anti-human IgG1 antibody conjugated with peroxidase (HRP) ( ) to detect bound antibodies.

[0034] Carry out as previously described (Williamso...

Embodiment 2

[0061] Defining the (linear or conformational) nature of the epitope recognized by the GRE1 monoclonal antibody

[0062] To define the (linear or conformational) nature of the epitope recognized by the GRE1 monoclonal antibody, Western Blot and Dot Blot were performed with both denatured and wild-type protein.

[0063] figure 2 Results of immunoblotting under denaturing conditions are shown. Proteins were denatured with β-mercaptoethanol (β-mer) or with sodium dodecyl sulfate (SDS). The commercial murine antibody was designated as Abcam, and the GRE1 anti-JCV VP1 monoclonal antibody was designated as GRE.

[0064] image 3 Results of dot blot experiments with both denatured VP1 and VP1 in wild-type conformation are shown. The commercial murine antibody was designated Abcam, while the GRE1 anti-JCV VP1 monoclonal antibody was designated IgG GRE.

[0065] The results showed that GRE1 was unable to bind to denatured forms of the protein, whereas it was only able to recogn...

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Abstract

The invention relates to a human neutralizing monoclonal antibody directed against the VP1 protein of JC virus, that is the virus responsible for progressive multifocal leukoencephalopathy (PML), as well as the use thereof in a therapeutic or prophylactic treatment of a JCV infection or of a disease associated with a JCV infection, such as pro¬ gressive multifocal leukoencephalopathy (PML), and the use thereof in the diagnosis of JCV infections or of diseases associated with JCV infections.

Description

technical field [0001] The present invention is within the field of immunology, in particular the field of neutralizing antibodies directed against antigens of viral pathogens. [0002] More specifically, the present invention relates to monoclonal antibodies directed against the VP1 protein of JC virus (JCV). Background technique [0003] JC virus (JCV) is a human polyomavirus of the family Polyomaviridae and is responsible for the extremely severe, often fatal demyelinating disease designated progressive multifocal Encephalopathy (progressive multifocal leukoencephalopathy, PML). JC virus has a non-enveloped icosahedral capsid that encapsulates a circular double-stranded DNA genome. The main component of the capsid is the viral protein VP1. Structural studies performed on virions revealed that the polyomavirus capsid is composed of 72 pentamers formed by VP1 monomers joined at the C-terminus. VP1 binds to receptors on target cells to initiate infection. [0004] JC vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08A61K39/42A61P31/20
CPCC07K2317/34C07K2317/33C07K16/084C07K2317/21C07K2317/30A61K2039/505C07K2317/76C07K2317/55A61P25/00A61P31/12A61P31/20Y02A50/30C07K16/081G01N33/56983G01N2469/10
Inventor 罗伯托·布廖尼马西莫·克莱门蒂
Owner POMONA RICERCA
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