MERS-CoV vaccine

A technology of immunogenicity and composition, applied in the vaccine field of Middle East Respiratory Syndrome Coronavirus, capable of solving limited and other problems

Active Publication Date: 2016-09-14
THE TRUSTEES OF THE UNIV OF PENNSYLVANIA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Human-to-human transmission of MERS-CoV

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] MERS-CoV shared spike antigen

[0180] As described elsewhere in this article, multiple strains of MERS-CoV include image 3 It is shown that a common spike antigen is present and generated to account for the differences between these multiple MERS-CoV strains. Therefore, the MERS-CoV shared spike antigen is derived from image 3 The corresponding spike antigens of the viruses listed in. Based on the spike antigen for germline analysis and relative to its component spike antigen ( image 3 , The mark "MERS-HCoV-shared" means MERS-CoV shared spike antigen) Place the MERS-CoV shared spike antigen. The MERS-CoV shared spike antigen also contains the N-terminal immunoglobulin E (IgE) leader sequence. The nucleic acid and amino acid sequences containing the MERS-CoV shared spike antigen are shown in Figure 8A with 8B in. in Figure 8A In, the underline indicates the nucleotide encoding the IgE leader sequence. in Figure 8B In the middle, the underline indicates the IgE lea...

Embodiment 2

[0184] MERS-CoV shared spike antigen lacking cytoplasmic domain

[0185] The spike antigen from the coronavirus is a type 1 membrane glycoprotein, which has a transmembrane domain, which in turn defines the cytoplasmic and non-cytoplasmic domains of the spike antigen. In order to determine the location of the cytoplasmic domain in the MERS-CoV spike antigen, web-based software was used to predict the location of the domain in various MERS-CoV spike antigens. Specifically, the web-based software Phobius and InterProScan were used in this analysis. Representative results from Phobius and InterProScan analysis are shown in Figure 5 with 6 in.

[0186] Analysis of this domain of the MERS-CoV spike antigen generates a second MERS-CoV common spike antigen, which lacks a cytoplasmic domain. Specifically, the nucleic acid sequence encoding the MERS-CoV shared spike antigen (described in Example 1, Figure 8A , SEQ ID NO: 1) was modified to insert two stop codons so that the translated p...

Embodiment 3

[0191] Expression and humoral response

[0192] Check the above-mentioned MERS-HCoV-WT and MERS-HCoV-ΔCD constructs to confirm that the corresponding antigens are expressed in cells and can be recognized by antibodies. The MERS-HCoV-WT and MERS-HCoV-ΔCD constructs were transfected into 293T cells along with pVAX1. pVAX1 served as a control.

[0193] Cell lysates were prepared two days after transfection and electrophoresed on a 5-15% SDS gel. Specifically, 10 μg, 25 μg, and 50 μg cell lysates were loaded into separate wells, and the cell lysates were obtained from 293T cells transfected with MERS-HCoV-WT or MERS-HCoV-ΔCD constructs. Sera from mice immunized with the MERS-HCoV-WT construct were then used for immunoblotting analysis. PVAX1( Picture 9 , No protein band was detected in the cell lysate obtained from the 293T cells transfected with pVAX1). For both the MERS-HCoV-WT and MERS-HCoV-ΔCD constructs, a protein band corresponding to the predicted molecular weight of the co...

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Abstract

Disclosed herein is a vaccine comprising a Middle East Respiratory Syndrome coronavirus (MERS-CoV) antigen. The antigen can be a consensus antigen. The consensus antigen can be a consensus spike antigen. Also disclosed herein is a method of treating a subject in need thereof, by administering the vaccine to the subject.

Description

[0001] Citation of related applications [0002] This application claims priority from U.S. Provisional Patent Application No. 61 / 910,153 filed on November 29, 2013, which application is hereby incorporated by reference. Technical field [0003] The present invention relates to a vaccine for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and a method of administering the vaccine. Background technique [0004] Coronavirus (CoV) is a common virus family all over the world and causes various diseases in humans from colds to severe acute respiratory syndrome (SARS). Coronavirus can also cause many diseases in animals. Human coronavirus 229E, OC43, NL63 and HKU1 are all endemic diseases in the human population. [0005] In 2012, a new type of coronavirus (nCoV) appeared in Saudi Arabia and is now called Middle East respiratory syndrome coronavirus (MERS-CoV) ( figure 1 ). MERS-CoV can be classified as a beta coronavirus ( figure 2 , Star-shaped MERS-CoV strain HCoV-EMC / 2012). ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/16A61K39/215A61K39/12
CPCA61K39/12A61K2039/53C12N2770/20034A61P31/14A61P37/04A61K39/215C12N7/00C07K14/005
Inventor 大卫·B·韦纳卡鲁皮亚·穆苏马尼尼兰詹·Y·萨尔德赛
Owner THE TRUSTEES OF THE UNIV OF PENNSYLVANIA
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