Genentech, Inc.

Washington Research Foundation

University of California

University of Washington

Benjamin D Hall

Gustav Ammerer

Ronald A Hitzeman

William J Rutter

Pablo D T Valenzuela

Franklin E Hagie Iv

Franklin E Hagie, Iv

Pablo D Valenzuela

The present invention relates to synthesis of HB Ag in yeast. Yeast expression vectors comprising a yeast promoter, ADH1, have been constructed. The region of the HBV genome coding for the S-protein, excluding a possible 163 amino acid presequence, has been transferred to the yeast expression vector. Using the described yeast vector, the successful synthesis of HB Ag by yeast has been achieved. The product is antigenic (reactive with anti-HB Ag), and a substantial portion is found associated with particles identical in electron microscopic appearance to those found in the serum of HBV-infected patients and in Alexander cells but having a smaller particle size diameter. The HB Ag synthesized by yeast has identical sedimentation behavior to purified, naturally occurring HB Ag particles purified from Alexander cells as measured by sucrose gradient sedimentation. The present invention demonstrates synthesis and assembly of a higher ordered multi-component structure resulting from expression of a heterologous DNA coding segment in a microorganism.

Synthesis of human virus antigens by yeast

The present invention relates to synthesis of HBsAg in yeast. Yeast expression vectors comprising a yeast promoter, ADH1, have been constructed. The region of the HBV genome coding for the S-protein, excluding a possible 163 amino acid presequence, has been transferred to the yeast expression vector. Using the described yeast vector, the successful synthesis of HBsAg by yeast has been achieved. The product is antigenic (reactive with anti-HBsAg), and a substantial portion is found associated with particles identical in electron microscopic appearance to those found in the serum of HBV-infected patients and in Alexander cells but having a smaller particle size diameter. The HBSAg synthesized by yeast has identical sedimentation behavior to purified, naturally-occurring HBsAq particles purified from Alexander cells as measured by sucrose gradient sedimentation. The present invention demonstrates synthesis and assembly of a higher ordered multi-component structure resulting from expression of a heterologous DNA coding segment in a microorganism.

DNA expression vectors capable, in a transformant strain of yeast, of expressing a polypeptide under the control of a genetically distinct yeast promoter, processes of forming transformant strains of yeast and transformed yeast strains are disclosed.

Expression of polypeptides in yeast

A DNA expression vector capable, in a transformant strain of yeast, of expressing a biologically competent polypeptide ordinarily exogenous to yeast under the control of a genetically distinct yeast promoter, the polypeptide not being required for the growth of the transformant; the process of forming the transformant strain of yeast; and the transformant strain of yeast.

The present invention relates to synthesis of HBsAg in yeast. Yeast expression vectors comprising a yeast promoter, ADHl, have been constructed. The region of the HBV genome coding for the S-protein, excluding a possible 163 amino acid presequence, has been transferred to the yeast expression vector. Using the described yeast vector, the successful synthesis of HBsAg by yeast has been achieved. The product is antigenic (reactive with anti-HBsAg), and a substantial portion is found associated with particles identical in electron microscopic appearance to those found in the serum of HBV-infected patients and in Alexander cells but having a smaller particle size diameter. The HBsAg synthesized by yeast has identical sedimentation behavior to purified, naturally-occurring HBsAg particles purified from Alexander cells as measured by sucrose gradient sedimentation. The present invention demonstrates synthesis and assembly of a higher ordered multi-component structure resulting from expression of a heterologous DNA coding segment in a microorganism.

Growing community of inventors

Bellevue, WA, United States of America

Benjamin D Hall