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Antibody-mediated neutralization of chikungunya virus

A Kungunya fever, antibody technology, applied in the fields of medicine, infectious diseases and immunology

Active Publication Date: 2018-02-16
VANDERBILT UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are no licensed vaccines or antiviral treatments to prevent or treat CHIKV infection

Method used

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  • Antibody-mediated neutralization of chikungunya virus
  • Antibody-mediated neutralization of chikungunya virus
  • Antibody-mediated neutralization of chikungunya virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0191] Example 1: Materials and methods

[0192] Isolation of human mAbs. PBMCs were obtained from persons ~5 years after documented symptomatic CHKV infection in Sri Lanka. B cells were transformed in 384-well plates with EBV in the presence of CpG. Supernatants from the resulting B-cell lymphoblastoid cell lines were screened for the presence of human CHKV-specific binding antibodies by ELISA using live CHIKV vaccine strain 181 / 25 virus as antigen. Transformed B cells are harvested and fused into myeloma cell lines, distributed into culture plates and expanded, and selected by growth in hypoxanthine-aminopterin-thymidine medium containing ouabain. Cloning of hybridomas by single cell sorting. Supernatants from clonal hybridomas grown in serum-free medium were collected, purified from clarified medium by protein G chromatography and concentrated.

[0193] Neutralization assay. Purified IgG mAb proteins were tested for neutralizing activity using CHKV viral replicon parti...

Embodiment 2

[0224] Example 2 - Results

[0225] Isolation of CHIKV-specific human mAbs. The inventor of the present invention acquired CHIKV infection in Sri Lanka from 2006 and developed fever, arthralgia and rash ( Figure 4 ) isolated a panel of mAbs from a single individual. The online approach provides the clinical course and B cell transformation and screening program. They transformed B cells in a single blood sample collected from a donor five and a half years after natural infection in two separate experiments. They observed a virus-specific B cell frequency of approximately 1 in 1,000 total B cells and established 30 stable hybridomas from B cell lines secreting antibodies that bind to the virus. The mAb panel contained multiple subclasses of IgG with 24 IgGl, 3 IgG2 and 2 IgG3; one was not identified due to poor hybridoma growth (Table 5).

[0226] Evaluation of mAb neutralization. Eighteen mAbs exhibited neutralizing activity against the Asian CHIKV strain SL15649-GFP vir...

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PUM

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Abstract

The present disclosure is directed to antibodies binding to and neutralizing Chikungunya virus (CHIKV) and methods for use thereof.

Description

[0001] This application claims the benefit of priority to US Provisional Application Serial No. 62 / 147,354, filed April 14, 2015, the entire contents of which are hereby incorporated by reference. [0002] background [0003] This invention was made with government support under grant numbers K08 AI103038, F32 AI096833 and U54AI057157 awarded by the National Institutes of Health. [0004] Background of the invention 1. Technical field [0005] The present disclosure relates generally to the fields of medicine, infectious disease, and immunology. More specifically, the present disclosure relates to antibodies that neutralize Chikungunya virus. The government has certain rights in this invention. 2. Background technology [0006] Chikungunya virus (CHIKV) is an enveloped positive-sense RNA virus in the genus Alphavirus of the Togaviridae family and is transmitted by the Aedes mosquito. Mature CHIKV virions contain two glycoproteins, E1 and E2, which are produced by proteo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/395C07K16/10C12N5/12C12N15/13
CPCC07K16/1081A61P31/14G01N33/577G01N33/56983C07K2317/21C07K2317/34C07K2317/565C07K2317/76C07K2317/92A61K2039/505G01N2333/181G01N2469/10A61K39/00C12Q1/70A61K39/395A61K2039/507Y02A50/30A61K39/12
Inventor J·E·克罗S·A·史密斯T·德莫迪L·席尔瓦
Owner VANDERBILT UNIV
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