Arrayed detector system for measurement of influenza immune response

Abstract

A sensor chip for detecting an immune response against an influenza virus, the sensor chip including a substrate having a surface and a plurality of hemagglutinin polypeptides bound to discrete locations on the surface of the substrate, each hemagglutinin polypeptide having a hemagglutinin epitope. Detection devices containing the sensor chip and methods of detecting influenza immune responses are also described herein.

1 .- 13 . (canceled) 
     
     
         14 . A method for screening efficacy of an influenza vaccine, said method comprising:
 administering an influenza vaccine to an individual;   obtaining a serum sample for the individual; and   sensing an anti-influenza antibody in the sample, wherein said sensing comprises:
 illuminating a surface of a multilayered substrate comprising hemagglutinin polypeptides from different influenza strains or isolates, wherein said hemagglutinin polypeptides comprise a hemagglutinin epitope and are covalently attached to the surface at different locations and retain conformational epitopes that are capable of binding to anti-influenza antibodies; 
 contacting the surface with the sample under conditions effective to allow an anti-influenza antibody in the sample to bind specifically to a hemagglutinin polypeptide recognized by the antibody; and 
 detecting light reflected from the surface under conditions effective to identify hemagglutinin polypeptides bound by an antibody of the sample to measure the anti-influenza immune response generated by the influenza vaccine. 
   
     
     
         15 . The method according to  claim 14 , wherein the multilayered substrate is an arrayed imaging reflectometry (AIR) sensor chip. 
     
     
         16 . The method according to  claim 14 , wherein said illuminating is carried out by directing a light at the surface. 
     
     
         17 . The method according to  claim 14 , wherein the sample is obtained from an individual selected from the group of humans and non-human primates, livestock, domesticated animals, and wild animals. 
     
     
         18 . The method according to  claim 14 , wherein the hemagglutinin polypeptides are full length hemagglutinin proteins. 
     
     
         19 . The method according to  claim 14 , wherein the hemagglutinin polypeptides are fragments of full length hemagglutinin proteins. 
     
     
         20 . The method according to  claim 19 , wherein the hemagglutinin polypeptide fragments each comprise an immunodominant hemagglutinin epitope of their respective native influenza strain. 
     
     
         21 . The method according to  claim 14 , wherein the hemagglutinin polypeptides are covalently attached to the substrate via a glutaraldehyde linker. 
     
     
         22 . The method according to  claim 14 , wherein each of said locations comprises a concentration of hemagglutinin polypeptide of about 100 fg/mm 2  to about 100 ng/mm 2 . 
     
     
         23 . The method according to  claim 14 , wherein the multilayered substrate further comprises:
 neuraminidase polypeptides from different influenza strains or isolates, wherein said neuraminidase polypeptides from each individual strain or isolate comprise a neuraminidase epitope and are covalently attached to the surface at different locations;   wherein said neuraminidase polypeptides retain conformational epitopes that are capable of binding to anti-influenza antibodies; and   wherein said surface with neuraminidase polypeptides covalently attached at the different locations forms a coating on the surface that results in destructive interference of polarized light illuminating said surface at an appropriate incident angle and wavelength in the absence of antibody binding.   
     
     
         24 . The method according to  claim 23 , wherein the neuraminidase polypeptides are full length neuraminidase proteins. 
     
     
         25 . The method according to  claim 23 , wherein the neuraminidase polypeptides are fragments of full length neuraminidase proteins. 
     
     
         26 . The method according to  claim 25 , wherein the neuraminidase polypeptide fragments each comprise an immunodominant neuraminidase epitope of their respective native influenza strain. 
     
     
         27 . The method according to  claim 14 , wherein said detecting is carried out with a detector that is positioned to detect light reflected from the surface to thereby determine whether an antibody binds to the hemagglutinin polypeptides, wherein a destructive interference of polarized light illuminating the surface occurs in an absence of antibody binding and antibody produces a changes in reflectance detectable by the detector.