Detection of antiviral antibodies with predetermined specificity using synthetic peptide--beta-lactamase conjugates: application to antibodies specific for the preS region of the hepatitis B virus envelope proteins.
Amino acid sequences coded for by the preS region of the hepatitis B virus (HBV) envelope gene are present both in HBV and in subviral hepatitis B surface antigen (HBsAg) particles. Consequently, anti-preS-specific antibodies are elicited during the course of HBV infection. Such antibodies are virus-neutralizing. Therefore, it is important to determine whether or not vaccination with HBsAg also induces an anti-preS-specific immune response. We describe here an enzyme-linked immunosorbent assay applicable for the screening of sera from vaccinated individuals for anti-preS antibodies. IgG from serum specimens was adsorbed to staphylococcal Protein A on a superparamagnetic support and subsequently mixed with a synthetic peptide analogue [preS(120-145)] covalently linked to beta-lactamase. The presence of anti-preS in serum specimens resulted in binding of the conjugated beta-lactamase to the magnetic support. The adsorbed enzyme was quantified colorimetrically.
A R
AR
Neurath
S B
SB
Kent
N
N
Strick
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes; D006508:Hepatitis Antibodies / Q000032:analysis* / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D009842:Oligopeptides / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology; D011863:Radioimmunoassay; D014759:Viral Envelope Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology; D001618:beta-Lactamases / Q000378:metabolism
D000939:Epitopes; D006508:Hepatitis Antibodies; D006514:Hepatitis B Surface Antigens; D009842:Oligopeptides; D011498:Protein Precursors; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; D001618:beta-Lactamases