Assessment of synthetic peptides for hepatitis A diagnosis using biosensor technology.
In the present work we demonstrate the application of a commercial biosensor instrument (BIACORE 1000, Biacore AB, Uppsala) for the detection of antibodies against the hepatitis A virus (HAV) in human serum samples using linear and branched synthetic peptides related to the VP3 capsid protein of HAV. We also studied the conformation of the synthetic peptides by circular dichroism (CD) in order to analyse the changes in secondary structure of the constructs that could influence their recognition by antibodies. Linear and dimeric VP3(110-121) multiple antigen peptides (MAP) were the most sensitive and appropriate for serological studies of serum from HAV infected patients using BIACORE. Immobilization of tetrameric MAPs via amine groups apparently failed to preserve the active conformation of the peptide epitope since it led to lower antibody binding compared to linear and dimeric peptides. The CD analysis showed that the tetrameric MAP constructs tend to adopt a beta-sheet structure due to intermolecular aggregation, which limits epitope accessibility. Our results demonstrate the value of biospecific interaction analysis technology using synthetic peptides for the diagnosis of acute hepatitis A.
Gómara
M J
MJ
Departament de Química de Pèptids i Proteïnes, IIQAB-CSIC, Jordi Girona 18-26, 08034, Barcelona, Spain.
Ercilla
G
G
Alsina
M A
MA
Haro
I
I
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques / Q000379:methods*; D002213:Capsid / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D036022:Capsid Proteins; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D006506:Hepatitis A / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D035922:Hepatitis A Antibodies; D006508:Hepatitis Antibodies / Q000097:blood*; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D036022:Capsid Proteins; D035922:Hepatitis A Antibodies; D006508:Hepatitis Antibodies; D009842:Oligopeptides; D010446:Peptide Fragments; C109000:VP3 peptide (110-121), hepatitis A virus