University System of Maryland

Daniel C Stein

Stefanie N Vogel

Philip R DeShong

Lenea Stocker

Katharina Richard

Charles K Stover

Robert T Yuan

Andrejez S Piekarowicz

The disclosure relates to compositions, pharmaceutical compositions, vaccines, and methods of making and using the same. The compositions of the disclosure are useful to stimulate an antigen-specific immune response and, in some embodiments, a protective immune response in an animal after challenge to pathogens. The compositions of the disclosure are also useful for treatment and prevention of disease in a subject such as cancer.

Compositions and vaccines comprising vesicles and methods of using the same

The invention relates to a hyperblebbing strain of,which produces large amounts of blebs useful for production of blebosomes containing antigens for use as a vaccine delivery vehicle or as a diagnostic reagent. The invention also relates to a method for producing high levels of a desired protein in purified form using the hyperblebbing strain of,, and to a vaccine delivery systems containing the blebosomes expressing the desired antigen.

Vaccine delivery system

E. coli bacterial strains encoding a restriction gene that degrades methylated DNA, and prevents cloning of genes expressing the methyltransferase responsible for methylation, are mutated by a chemical or physical mutagen, so as to make the restriction enzyme temperature sensitive. Mutant cells are rendered competent, and plasmids expected or known to contain genes encoding methyltransferase enzymes are introduced. The transformants grow at the permissive temperature, where the restriction enzyme system is inactivated due to the mutated gene. Successful clones, expressing a methyltransferase, can be quickly identified by those which grow at the permissive temperature, but not at the non-permissive temperature. The valuable methyltransferases, as well as restriction enzymes associated therewith, can accordingly be recovered in large quantity.

Mutant strain of E. coli for detection of methyltransferase clones

Growing community of inventors

Silver Spring, MD, United States of America

Daniel C Stein