Systems and methods for yeast vaccines are presented that allow for selection of tumor neoepitopes that are then used to generate a recombinant polytope for yeast expression with enhanced immunogenicity.

A recombinant protein comprising a functional polypeptide and, linked to the N-terminus of said functional polypeptide, an N-terminal spacer having a length such that the number of amino acid residues between a signal peptide cleaving site and an N-terminus proximal structural unit of said functional polypeptide is 14-24.

An expression system and process for the production of Diphtheria toxin polypeptides or mutated forms thereof, such as the toxoid CRM197 polypeptide, in genetically-modified E. coli with high yield is described. The system and process is based on the uncoupling of biomass growth from recombinant protein induction, i.e. using an inducer of protein production that cannot be used as a carbon source for growth by the bacteria. The use of specific components and conditions that improve protein yields are also described.

The present invention is a DNA expression vector comprising: a toxP; a mutant toxO that blocks Fe-mediated regulation of gene expression; and a DNA sequence encoding a protein, wherein the toxP and the mutant toxO regulate expression of the DNA segment encoding the protein. It is preferred that DNA expression vectors of the present invention include DNA sequences encoding a signal peptide so that a protein expressed is attached to the signal peptide prior to processing. Novel proteins are produced off of the DNA expression vector of the present invention.

The invention relates to proteins containing any of the sequences according to general formula (Ih):

X.sub.1CRX.sub.2X.sub.3X.sub.4X.sub.5(Ih)

where
X.sub.1 is F, Y, L, P, Q, M, V, W, A, T and
X.sub.2 is A, G, S, T and
X.sub.3 is V, A, I, L, M, D, H, S and
X.sub.4 is K, I, Q, R, H, S, F, M, N, L, V and
X.sub.5 is R, V, I, K, M, Q, E, F, L, N, Y, D, S, H. and
its salts, esters and pharmaceutically acceptable prodrugs. The invention further relates to pharmaceutical preparations and kits containing said proteins, to screening procedures using said proteins. The proteins of the invention are useful in the treatment or prevention of diseases that can be treated by inhibiting the complement system.

Provided is a protein complex including a botulinum toxin translocation domain and endolysin. When used, the protein complex including a botulinum translocation domain and endolysin according to the present invention exhibits an antibacterial effect and thus can be used as an antibacterial composition or an antibiotic.

The present invention is directed to a bioconjugate vaccine, such as an O 1-bioconjugate vaccine, comprising: a protein carrier comprising a protein carrier containing at least one consensus sequence, DIE-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; at least one antigenic polysaccharide from at least one pathogenic bacterium, linked to the protein carrier; and, optionally, an adjuvant. In another aspect, the present invention is directed to a method of producing an O 1-bioconjugate in a bioreactor comprising a number steps.

A fusion polypeptide, comprising an inclusion body tag fused to at least one polypeptide of interest, is provided. The inclusion body tag comprises an inclusion body forming (IBF) amino acid sequence selected from X.sub.14X.sub.15AX.sub.17X.sub.18X.sub.19GLTVA GMLG and sequences having at least 71% identity thereto. Also provided are inclusion bodies comprising the fusion polypeptide, as well as nucleic acids, expression vectors, host cells and methods for its production.

The present invention is a DNA expression vector comprising: a toxP: a mutant toxO that blocks Fe-mediated regulation of gene expression; and a DNA sequence encoding a protein, wherein the toxP and the mutant toxO regulate expression of the DNA segment encoding the protein. It is preferred that DNA expression vectors of the present invention include DNA sequences encoding a signal peptide so that a protein expressed is attached to the signal peptide prior to processing. Novel proteins are produced off of the DNA expression vector of the present invention.

The present invention resides in a method for the manufacture of a disulphide-requiring biopharmaceutical having an element of at least tertiary structure using wild type E. coli.