Whitehead Institute for Biomedical Research

Harvard College

Massachusetts Institute of Technology

The General Hospital Corporation

The Brigham and Women's Hospital, Inc.

Children's Medical Center Corporation

The General Hospital Corporation D/b/a Massachusetts General Hospital

Hidde L Ploegh

J Christopher Love

Jehnna Ronan

Harvey F Lodish

Jiahai Shi

Harold A Chapman

Hsiang-Ying Lee

Lenka Hoffman

Paula R Bryant

Matthew S Bogyo

Mohammad Rashidian

Lee Kim Swee

Novalia Pishesha

Richard J Riese

Daniel Griffith Anderson

Ralph Weissleder

Omar Khan

Tyler E Jacks

Edmund J Keliher

Richard O Hynes

David A Canner

Nicholas C Yoder

Noor Jailkhani

Jasdave S Chahal

Martin D Witte

Robert Samuel Langer

Guillermo A Ameer

Thomas J Gardella

Carla Guimaraes

Neil Vasdev

Huan Steven Liang

Robert S Langer

Maria Ann Rupnick

Ross W Cheloha

Yushu Joy Xie

Eric Grovender

Rober S Langer, Jr

Yushu J Xie

Maximilian Popp

Juanjo Cragnolini

John M Antos

Maximilian Wei-Lin Popp

Lenka Kundrat

Methods for developing disease-related nanobodies and related products and kits are provided. The disease-specific proteins are extracellular matrix (ECM) proteins, domains or epitopes that are associated with various aspects of disease and are not present, or are present in very low quantities, in non-diseased individuals. Highly effective nanobodies capable of specifically binding to these ECM protein epitopes useful in in vivo imaging assays, the detection, diagnosis and treatment of diseases as well as monitoring therapeutic progress in a patient with a disease are provided herein.

Nanobody based imaging and targeting of ECM in disease and development

Compositions and methods for modified dendrimer nanoparticle ('MDNP') delivery of therapeutic, prophylactic and/or diagnostic agent such as large repRNA molecules to the cells of a subject have been developed. MDNPs efficiently drive proliferation of antigen-specific T cells against intracellular antigen, and potentiate antigen-specific antibody responses. MDNPs can be multiplexed to deliver two or more different repRNAs to modify expression kinetics of encoded antigens and to simultaneous deliver repRNAs and mRNAs including the same UTR elements that promote expression of encoded antigens.

Compositions and methods for modified dendrimer nanoparticle delivery

Non-genetically engineered mammalian cells modified by sortase-mediated conjugation of an agent thereto are provided. Methods of conjugating agents to non-genetically engineered mammalian cells using sortase are provided. Methods of using the cells, e.g., for diagnostic and/or therapeutic purposes, are provided.

Protein modification of living cells using sortase

Provided herein are methods for producing site specific PEG modifications to single domain antibodies (e.g., VHHs). Methods for producing site-specifically conjugated bivalent single domain antibodies (e.g., VHHs) are also provided. Methods for labeling (e.g., with a fluorophore or radionuclide) site-specifically PEGylated single domain antibodies and site-specifically conjugated bivalent single domain antibodies are also provided.

Labeling of antibodies

Methods for the in vitro production of enucleated red blood cells and the enucleated red blood cells thus prepared are provided. Such enucleated red blood cells may express a sortaggable surface protein, which allows for surface modification in the presence of a sortase. Also described herein are surface modified enucleated red blood cells, e.g., conjugated with an agent of interest such as a peptide, a detectable label, or a chemotherapeutic agent, and uses thereof in delivering the agent to a subject.

In vitro production of red blood cells with proteins comprising sortase recognition motifs

Described herein are engineered ligand that binds a cell surface receptor (e.g., GPCR), with improved affinity, potency, and specificity. By conjugating a sub-optimal ligand for a cell surface receptor (e.g., GPCR) to a targeting molecule that binds an epitope (natural or exogenous epitope) in the extracellular portion of the cell surface receptor (e.g., GPCR), the affinity, potency, and/or specificity of the sub-optimal ligand is enhanced.

Engineered ligands and uses thereof

The present invention, in some aspects, provides methods, reagents, compositions, and kits for the radiolabeling of proteins, for example, of proteins useful for positron emission tomography (PET) or single-photon emission computed tomography (SPECT) (e.g., for diagnostic and therapeutic applications), using sortase-mediated transpeptidation reactions. Some aspects of this invention provide methods for the conjugation of an agent, for example, a radioactive agent or molecule to diagnostic or therapeutic peptides or proteins. Compositions comprising sortagged, radiolabeled proteins as well as reagents for generating radiolabeled proteins are also provided. Kits comprising reagents useful for the generation of radiolabeled proteins are provided, as are precursor proteins that comprise a sortase recognition motif.

18F labeling of proteins using sortases

In vitro production of red blood cells with sortaggable proteins

Screening assays and methods of performing such assays are provided. In certain examples, the assays and methods may be designed to determine whether or not two or more species can associate with each other. In some examples, the assays and methods may be used to determine if a known antigen binds to an unknown monoclonal antibody.

Screening assays and methods

Methods and reagents for the installation of click chemistry handles on target proteins are provided, as well as modified proteins comprising click chemistry handles. Further, chimeric proteins, for example, bi-specific antibodies, that comprise two proteins conjugated via click chemistry, as well as methods for their generation and use are disclosed herein.

Using sortases to install click chemistry handles for protein ligation

The present invention, in some aspects, provides methods, reagents, compositions, and kits for the radiolabeling of proteins, for example, of proteins useful for positron emission tomography (PET) or single-photon emission computed tomography (SPECT) (e.g., for diagnostic and therapeutic applications), using sortase-mediated transpeptidation reactions. Some aspects of this invention provide methods for the conjugation of an agent, for example, a radioactive agent or molecule to diagnostic or therapeutic peptides or proteins. Compositions comprising sortagged, radiolabeled proteins as well as reagents for generating radiolalebed proteins are also provided. Kits comprising reagents useful for the generation of radiolabeled proteins are provided, as are precursor proteins that comprise a sortase recognition motif.

F labeling of proteins using sortases

Growing community of inventors

Brookline, MA, United States of America

Hidde L Ploegh