Wisconsin Alumni Research Foundation

The Medical College of Wisconsin, Inc.

Mcw Research Foundation, Inc.

Cellsnap LLC

Joseph T Barbieri

Eric Arthur Johnson

Marite Bradshaw

William Howard Tepp

Sabine Pellett

Michael Baldwin

Dara W Frank

Sheng Chen

Amanda Przedpelski

Christina Pier

Arthur L Haas

Brian D Halligan

Scott M Kulich

Lawrence M Ryan

Daniel J McCarty

Ikuko Masuda

Christina L Pier

William H Tepp

Provided herein are antigenic peptides comprising the SARS-COV-2 spike protein receptor binding domain (CRBD) polypeptide or portions thereof, linked to a non-catalytic, non-toxic tetanus toxin variant (i.e., a modified tetanus toxin or 'MTT') and vaccine compositions comprising the same. In addition, provided herein are methods for making and using CRBD-MTT fusion proteins as immunogenic agents.

COVID-19 vaccine

Provided herein are engineered non-catalytic, non-toxic tetanus toxin variants and methods of using such engineered tetanus toxin variants as low dose, protective vaccines that are non-toxic and more potent than their respective chemically inactivated toxoids. In addition, provided herein are conjugate vaccine carriers comprising engineered tetanus toxin variants and methods of using such conjugate vaccines to elicit T-cell dependent immune memory responses which can target a broad spectrum of microbial pathogens as a single vaccine.

Modified clostridial neurotoxins as vaccines and conjugate vaccine platforms

The present disclosure relates to chimeric neurotoxins and uses thereof. In particular, provided herein are chimeric botulinum neurotoxins with altered properties and uses thereof (e.g., research, screening, and therapeutic uses).

Hybrid neurotoxins and uses thereof

The present invention provides a novel modified BoNT/E catalytic domain and methods of use thereof. In one embodiment, the light chain residue 224, or a residue corresponding to residue 224, of the modified BoNT/E catalytic domain has been altered to be aspartic acid or glutamic acid. The modified catalytic domain cleaves SNAP23 but does not cleave SNAP29 or SNAP47, providing novel methods of treating diseases including without limitation, asthma, CF, chronic obstructive pulmonary, gastric acid efflux and inflammation, immune disorders with a cytokine component or cancers with a cytokine component.

Engineered botulinum neurotoxin

The present invention provides a preparation of botulinum toxin light chain type A or E, wherein the preparation is both catalytically active and soluble. Preferably, the preparation consists essentially of amino acid residues 1 through 425 of the botulinum toxin light chain type A. A method of screening inhibitors is also provided, wherein the method comprises exposing a test inhibitor to the preparation of botulinum toxin light chain type A and evaluating the biological activity of the preparation. In another embodiment, a method of providing a catalytically active, soluble preparation of botulinum toxin light chain, type A is provided, wherein the method comprises obtaining an expression vector comprising a DNA sequence encoding amino acid residues 1-425 and expressing a polypeptide.

Method of preparing botulinum neurotoxin type E light chain

A method of producing botulinum toxin C-terminal receptor binding domain (HCR) is disclosed. The one embodiment, the method comprises the steps of (a) preparingtransformed with an expression vector comprising DNA encoding HCR protein, (b) inducing expression of the HCR protein at a reduced temperature in a culture media, and (c) purifying the HCR protein via extraction, wherein the extraction comprises a clarification by centrifugation and a filtration, wherein the purified HCR protein is at least 10 mg/L of culture medium.

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Method for preparing botulinum neurotoxin type A light chain

We have cloned and sequenced the cDNA encoding the 61 kD active fragment of a unique porcine chondrocyte nucleotide pyrophosphohydrolase (NTPPHase) from a porcine chondrocyte library. Degenerate oligonucleotides, corresponding to the N-terminal amino acid sequence of this peptide were hybridized to porcine chondrocyte cDNA and used to amplify DNA encoding the N-terminal sequence of 61 kD with the polymerase chain reaction (PCR). The PCR products were then used as probes to clone the entire open reading-frame for the 61 kD fragment from a porcine chondrocyte cDNA library. The length of the cloned cDNA was 2509 bp. Translation of the open-reading-frame predicts the 61 kD fragment to be a 459 amino acid protein. BLAST and FASTA analysis confirmed that this amino acid sequence was unique and did not possess high homology to any known proteins in the non-redundant protein data base. Limited homology (17%) between the 61 kD fragment and several prokaryotic and eukaryotic ATP pyrophosphate-lyase (adenylate cyclase) was detected. Northern blot analysis of porcine chondrocyte RNA showed that the DNA encoding the 61 kD fragment hybridized to a 4.3 kbp RNA transcript. Human chondrocyte RNA also hybridized to this porcine DNA probe. Coupled in vitro transcription translation of an expression vector containing the DNA insert in frame showed the expression of a 61 kD protein.

Cloned nucleotide pyrophosphohydrolase and uses thereof

A genetic construct containing a coding region for exoenzyme S activity from Pseudomonas aeruginosa is disclosed. A essentially pure protein preparation of the 49 kDa form of exoenzyme S is also disclosed. The protein product of the genetic construct may be used to modify the RAS protein function in mammalian carcinomas, used as a vaccine, or used to diagnose Pseudomonas aeruginosa infection.

Growing community of inventors

New Berlin, WI, United States of America

Joseph T Barbieri