Identification of Additional Antigenic Sites on Dane Particles and the Tubular Forms of Hepatitis B Surface Antigen

Abstract

SUMMARYAdditional antigenic sites, distinct from those present on spherical 20 nm diam. particles of hepatitis B surface antigen (HB~Ag), are exposed on the surface of Dane particles and tubular forms of HB~Ag. The immunological relationship of these sites to e-antigen, an antigen detected earlier in HB~Ag-positive sera from patients with chronic hepatitis, cirrhosis or acute hepatitis but not in healthy HB~Ag-carriers, was established by immune electron microscopy and affinity chromatography. These findings suggest that e-antigen may be potentially useful in active immunization against hepatitis B.

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

“…1. Results of ELISA tests with fivefold serial dilutions of rabbit antisera to HBV (O) (Neurath et al, 1976(Neurath et al, , 1985a, and to the synthetic peptide preS(120 145) (•) (Neurath et al, 1984a), and with dilutions of a serum pool from recipients of a hepatitis B vaccine (Iq) (McAuliffe et al, 1982). The enzyme-labelled antigen was preS(120-145)-/Mactamase.…”

“…Additional sera were used in the course of the development of the ELISA test: rabbit antisera to HBV from which antibodies Io the S-protein had been removed (Neurath et al, 1976: rabbit antisera to the synthetic peptides preS(120-145) and preS(12 32) (Neurath et al, 1984a~ 1985a and pooled serum from individuals vaccinated with an experimental vaccine (McAuliffe et al, 1982). The latter serum pool contained sera of individuals positive for antibodies with anti-preS specificity as determined by double antibody radioimmunoassays (RIA; Neurath et al, 1985a).…”

Hepatitis B virus vaccine: identification of HBsAg/a and HBsAg/d but not HBsAg/y subtype antigenic determinants on a synthetic immunogenic peptide.

“…Such vaccines could contain unwanted antigenic components, derived from elsewhere in the virion or its associated particles, or from the host cells in which these were synthesized. It is difficult, ifnot impossible, to remove all detectable host components-e.g., human serum albumin, IgG, and some other serum proteins-from even the most highly purified HBsAg particles (45)(46)(47)(48). Indeed, when such particles derive from HBV e antigen-positive donors, they adsorb efficiently to aggregated human serum albumin (48).…”

Expression in Escherichia coli of a cloned DNA sequence encoding the pre-S2 region of hepatitis B virus.

“…This fusion protein, which comprised as much as 3% of the total bacterial protein, was purified to >90% homogeneity by affinity chromatography on p-aminophenyl-.3D-thiogalactoside-Sepharose. of components detected in earlier studies on the protein composition of the HBV envelope and of HBsAg.Although the pre-Sl and pre-S2 sequences have a relatively low abundance in HBV or HBsAg isolated from the serum of HBV carriers, their biological role has already been explored by using appropriate polyclonal or monoclonal antibodies against HI3V (7,8), synthetic peptides (9), and proteins synthesized in cells transfected with expression vector containing the total S region or large portions of pre-S (10, 11).For example, the amino acid sequences, encoded in the pre-S1and pre-S2 regions have been shown to be involved in specific binding of HBV (HBsAg) to the surface of human hepatocytes (12). The amino acid sequence ofpre-S2 has also been found to bind human and chimpanzee glutaraldehyde crosslinked polyalbumin (11,13,14).…”

“…Although the pre-Sl and pre-S2 sequences have a relatively low abundance in HBV or HBsAg isolated from the serum of HBV carriers, their biological role has already been explored by using appropriate polyclonal or monoclonal antibodies against HI3V (7,8), synthetic peptides (9), and proteins synthesized in cells transfected with expression vector containing the total S region or large portions of pre-S (10, 11).…”