title: Mimotope ELISA for detection of broad spectrum antibody against avian H5N1 influenza virus.
abstract: BACKGROUND
  We have raised a panel of broad spectrum neutralizing monoclonal antibodies against the highly pathogenic H5N1 avian influenza virus, which neutralize the infectivity of, and afford protection against infection by, most of the major genetic groups of the virus evolved since 1997. Peptide mimics reactive with one of these broad spectrum H5N1 neutralizing antibodies, 8H5, were identified from random phage display libraries.
  
  
  METHODS
  The amino acid residues of the most reactive 12mer peptide, p125 (DTPLTTAALRLV), were randomly substituted to improve its mimicry of the natural 8H5 epitope.
  
  
  RESULTS
  133 reactive peptides with unique amino acid sequences were identified from 5 sub-libraries of p125. Four residues (2,4,5.9) of the parental peptide were preserved among all the derived peptides and probably essential for 8H5 binding. These are interspersed among four other residues (1,3,8,10), which exhibit restricted substitution and probably could contribute to binding, and another four (6,7,11,12) which could be randomly substituted and probably are not essential for binding. One peptide, V-1b, derived by substituting 5 of the latter residues is the most reactive and has a binding constant of 3.16×10(-9) M, which is 38 fold higher than the affinity of the parental p125. Immunoassay produced with this peptide is specifically reactive with 8H5 but not also the other related broad spectrum H5N1 avian influenza virus neutralizing antibodies. Serum samples from 29 chickens infected with H5N1 avian influenza virus gave a positive result by this assay and those from 12 uninfected animals gave a negative test result.
  
  
  CONCLUSIONS
  The immunoassay produced with the 12 mer peptide,V1-b, is specific for the natural 8H5 epitope and can be used for detection of antibody against the broad spectrum neutralization site of H5N1 avian influenza virus.
authors.affiliation: National Institute of Diagnostics and Vaccine Development in Infectious Diseases, The Key Laboratory of Education Minister for Cell Biology and Tumor Cell Engineering of Xiamen University, School of Life Sciences, Xiamen University, Xiamen, China.
authors.forename: Yingwei
authors.initials: Y
authors.lastname: Chen
authors.forename: Wenxin
authors.initials: W
authors.lastname: Luo
authors.forename: Huijuan
authors.initials: H
authors.lastname: Song
authors.forename: Boyuan
authors.initials: B
authors.lastname: Yin
authors.forename: Jixian
authors.initials: J
authors.lastname: Tang
authors.forename: Yixin
authors.initials: Y
authors.lastname: Chen
authors.forename: Mun Hon
authors.initials: MH
authors.lastname: Ng
authors.forename: Anthony E T
authors.initials: AE
authors.lastname: Yeo
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Ningshao
authors.initials: N
authors.lastname: Xia
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D057134:Antibodies, Neutralizing / Q000032:analysis* / Q000276:immunology*; D000918:Antibody Specificity; D040761:Biomimetic Materials / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology*; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000939:Epitopes; D009842:Oligopeptides