Method for detection of mers-cov with igy antibodies

Abstract

A method for detecting MERS-CoV at high sensitivity and specificity using IgY antibodies that bind to MERS-CoV N protein, its fragments and domains. Isolated or purified IgY monospecific antibodies to MERS-CoV N protein.

1 . A method for detecting MERS-CoV in a sample, comprising
 contacting the sample with a monospecific IgY antibody that recognizes MERS-CoV nucleocapsid protein (“N protein”) and detecting complex formation or binding of the IgY antibody to the MERS-CoV.   
     
     
         2 . The method of  claim 1 , which is an ELISA sandwich assay, wherein the IgY antibody that recognizes MERS-CoV N protein is a capturing antibody bound to a substrate, wherein the capturing antibody bound to the substrate is contacted with the sample suspected of containing MERS-CoV, and wherein complex formation is measured by the binding of a tagged secondary antibody that recognizes MERS-CoV. 
     
     
         3 . The method of  claim 1  that is an immunoswab assay, wherein said contacting comprises contacting the sample with IgY antibodies bound to a swab, removing unbound sample, and detecting complex formation. 
     
     
         4 . The method of  claim 1 , wherein said sample contains no more than 20 ng/ml of MERS-CoV N protein. 
     
     
         5 . The method of  claim 1 , wherein the IgY antibody does not substantially interact with rheumatoid factor (RF), human anti-mouse IgG antibodies (HAMA), complement components and/or mammalian Fc receptors. 
     
     
         6 . The method of  claim 1 , wherein the IgY antibody is a polyclonal antibody produced by immunizing chickens with isolated or recombinant MERS-CoV N protein and recovering IgY antibody from eggs laid by the immunized chickens. 
     
     
         7 . The method of  claim 1 , wherein the IgY antibody comprises a polyclonal monospecific IgY antibody for MERS-CoV N protein. 
     
     
         8 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to a peptide epitope of MERS-CoV N protein that ranges in length from 6 to 20 amino acids of the amino acid sequence described by SEQ ID NO: 1. 
     
     
         9 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to an N-terminal domain segment of MERS CoV N protein comprising residues 1-36 of SEQ ID NO: 1. 
     
     
         10 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to an N-terminal domain segment of MERS CoV N protein comprising residues 37-164 of SEQ ID NO: 1. 
     
     
         11 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to a central segment of MERS CoV N protein comprising residues 165-238 of SEQ ID NO: 1; or that binds to a disordered region or LKR containing a stretch of serine and arginine residues and comprising residues 156 to 206 of SEQ ID NO: 1 or 
       
         
           
                 
               
                   (SEQ ID NO: 3) 
                 
                   LPKNFHIEGTGGNSQSSSRASSVSRNSSRSSSQGSRSGNSTRGTSPGPSG 
                 
                     
                 
                   I. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         12 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to a C-terminal domain segment of MERS CoV N protein comprising residues 239-362 of SEQ ID NO: 1. 
     
     
         13 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to a C-terminal domain segment of MERS CoV N protein comprising residues 363-413 of SEQ ID NO: 1. 
     
     
         14 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to one or more residues in a peptide segment of SEQ ID NO: 1 selected from the group comprising g residues 4-12, 42-47, 50-58, 60-66, 92-102, 108-126, 144-150, 172-180, 206-225, 230-240, 261-268, 296-304, 324-337, 347-353and 399-405 described by SEQ ID NO: 1. 
     
     
         15 . The method of  claim 1 , wherein the IgY antibody comprises a monospecific antibody that binds to one or more residues in a peptide segment of SEQ ID NO: 1 selected from the group comprising:
 K346, W347, L348, and E349;   Y327, F328, L329, and R330;   L350, L351, E352, O353, N354, 1355, D356, A357, Y358, K359, T360, F361, P362, K363, K364, and E365;   D320, D321, H322, G323, N324, P325, and V326;   H252, K253, R254, T255, S256, T257, K258, S259, F260, N261 , M262, V263, Q264, A265, F266, G267, L268, R269, G270, P271, G272, D273, L274, Q275, G276, N277, F278, G279, D280, L281, Q282, L283, N284, K285, L286, G287, T288, E289, D290, P291, and R292;   R399, P400, S401, V402, Q403, P404, G405, P406, M407, 1408, D409, V410, N411, T412, and D413;   W293, P294, 0295, 1296, A297, and E298;   F312, K313, L314, T315, H316, Q317, N318, and N319; and   L299, A300, P301, T302, A303, 5304, A305, F306, M307, G308, M309, S310, and Q311.   
     
     
         16 . The method of  claim 1 , wherein the IgY antibody comprises a mixture of two or more monospecific IgY antibodies to MERS CoV N protein epitopes or domains. 
     
     
         17 . An antibody conjugate comprising a monospecific polyclonal IgY antibody that binds to MERS-CoV N protein that is conjugated to at least one detectable marker or to a therapeutic agent. 
     
     
         18 . The antibody conjugate of  claim 17  that binds to the N terminal domain segment MERS-CoV N protein comprising residues 37-164 of SEQ ID NO: 1. 
     
     
         19 . The antibody conjugate of  claim 17  comprising a monospecific polyclonal IgY antibody that binds to the C terminal domain segment of MERS-CoV comprising residues 239-362 of SEQ ID NO: 1. 
     
     
         20 . A method for treating a subject at risk of being infected by, or infected by, MERS-CoV comprising administering an IgY antibody that binds to MERS-CoV N protein to the subject.