Production of a soluble and secreted antigenic fragment of HBsAg in yeast

• Published in: Biochemical and biophysical research communications Vol. 184; no. 1; pp. 50 - 59
• Main Authors: Baba, Elio Hideo, Berkower, Ira
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.04.1992; Elsevier
• Subjects: Amino Acid Sequence; Antigenic determinants, haptens, artificial antigens; Antigens; Base Sequence; Biological and medical sciences; DNA, Viral - genetics; DNA, Viral - isolation & purification; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genetic Vectors; Hepatitis B Surface Antigens - biosynthesis; Hepatitis B Surface Antigens - genetics; Hepatitis B Surface Antigens - isolation & purification; Hepatitis B virus - genetics; Mating Factor; Molecular immunology; Molecular Sequence Data; Oligodeoxyribonucleotides - chemical synthesis; Peptide Biosynthesis; Peptide Fragments - biosynthesis; Peptide Fragments - genetics; Peptide Fragments - isolation & purification; Peptides - genetics; Peptides - isolation & purification; Pheromones - genetics; Plasmids; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - isolation & purification; Restriction Mapping; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Transformation, Genetic; yeast alpha mating factor; hepatitis B virus surface antigen; human major histocompatibility complex; fragment 1–60 of HBsAg; Fungi; Hepatitis B surface antigen; Yeast; Antigenicity; Antigenic determinant; Peptide fragment; Ascomycetes; Saccharomyces cerevisiae; Thallophyta
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(92)91156-K

We have produced a fragment of hepatitis B surface antigen (HBsAg) corresponding to amino acids 1–60 as a fusion protein with the alpha mating factor of yeast. The product is secreted from yeast as a soluble monomer that expresses HBsAg antigenicity. Unlike other heterologous fusion proteins, it is not processed by the Lys-Arg endoprotease, possibly due to a proline in the linker between the two coding sequences. The resulting soluble fragment will enable us to map the immunodominant sites of HBsAg recognized by T cells and to identify additional factors contributing to vaccine potency.