ABSTRACT
HBsAg containing pre-s (2), purified by affinity chromatography with PHSA coupled to CNBr-activated sepharose 4B, was used as immunogen to immunize in vivo spleen of BALB/C female mice. The spleen cells from immunized mice were fused with mouse myeloma cells (SP2/0-Ag). Clones producing the antibody were screened by serial enzyme immunosorbent assay, and subclone cultures were isolated by limiting dilution. Finally, three clones, which were positive for anti-pre-s (2) antibody, were selected. The clones were still stable by several passages. They were injected i. p. to prime-treated BALB/C mice to produce ascitic fluid. The liters of anti-pre-s (2) antibodies in the ascitic fluid were above 105.Isotype analysis revealed that all immunoglobulins were of IgGl subclass of mouse. The purified McAb for pre-s (2) conjugated with horseradish peroxidase were used to develop a sandwich-type assay (ELISA). Using the assay, pre-s (2), the receptor for PHSA,was detected in serum of the patients with HBV infection. The assay method was more sensitive, specific, reproducible and reliable. The assay may be practical in the routine work of hepatitis B immunodiagnosis, and worth further clinical trial.
ABSTRACT
Pre-S2 protein in 78 samples of liver from patients with various types of hepatitis B was determined by immunohistochemical method.45 out of 78 samples (57.69%) were positive for pre-S2.The location and expression patterns of pre-S2 were similar to those of HBsAg,i.e.,pre-S2 was found in cytoplasm of hepatocytes,epithelial cells of bile ducts,and hepatocellular carcinoma cells,etc,Four different patterns diffuse,inclusion,submembranous,and membranous type were demonstrated.The membranous expression of pre-S2 was often associated with activity of liver diseases.The positive rate of pre-S2 was significantly higher in HBcAg positive group than HBcAg negative group.It seems that pre-S2 protein of HBV may be a replicative marker of HBV.