Single-domain VH H antibodies directed to norovirus GI.1 and GII.4 and their use
Inventors
Bok, Karin • Garaicoechea, Lorena Laura • Parreno, Viviana • Aguilar, Andrea Pamela • Bok, Marina • Green, Lisbeth Kim • Sosnovtsev, Stanislav Vladimirovich
Assignees
Instituto Nacional de Tecnologia Agropecuaria INTA • US Department of Health and Human Services
Publication Number
US-11325965-B2
Publication Date
2022-05-10
Expiration Date
2034-05-09
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Abstract
Isolated VHH monoclonal antibodies are disclosed that specifically bind to a Norovirus polypeptide. In some embodiments, the Norovirus is a Genogroup I Norovirus or a Genogroup II Norovirus. In other embodiments, the Norovirus is Norwalk or MD2004 virus. In some embodiments, the monoclonal antibodies specifically bind VP1. Also disclosed are compositions including the disclosed antibodies, nucleic acids encoding these antibodies, expression vectors including the nucleic acids, and isolated host cells that express the nucleic acids. The antibodies and compositions disclosed herein can be used for detecting the presence of a Norovirus in a biological sample, or detecting a Norovirus infection. Also disclosed are methods of treating and/or preventing a NoV infection.
Core Innovation
The invention provides isolated VHH monoclonal antibodies, also known as camelid nanobodies, that specifically bind to Norovirus (NoV) polypeptides, such as the major capsid protein VP1. These antibodies can specifically target NoVs from Genogroup I or Genogroup II, including strains like Norwalk virus (GI.1) and MD2004 virus (GII.4). The antibodies include heavy chain variable domains comprising complementarity determining regions (CDRs), with detailed sequences provided (SEQ ID NOs: 1-30). The VHH antibodies can be llama-derived, humanized, or chimeric monoclonal antibodies and can be produced recombinantly.
The problem being addressed is the lack of effective reagents for detecting and treating Norovirus infections, which are a leading cause of epidemic gastroenteritis and associated with significant morbidity and mortality, particularly in children, the elderly, and immunocompromised individuals. Prior to this invention, there were no available vaccines or therapies for NoV diarrhea, partly due to the absence of an in vitro cell culture system for NoV and the need for antigenically relevant neutralizing antibodies.
The invention also discloses compositions that include the isolated antibodies or antigen binding fragments, nucleic acids encoding these antibodies, expression vectors, and host cells expressing the antibodies. Methods for detecting NoV in biological or environmental samples using the antibodies are provided. These methods comprise contacting a sample with the VHH monoclonal antibody or antigen binding fragment and detecting binding, thereby indicating the presence of the virus. Therapeutic methods using the antibodies or nucleic acids encoding them to treat or prevent NoV infections in subjects are also encompassed, with examples including administration to immunocompromised individuals or children.
Claims Coverage
The patent contains multiple inventive features focused on isolated VHH monoclonal antibodies specific to Norovirus polypeptides, their use in detection methods, and compositions for treatment.
Unique VHH monoclonal antibodies specifically binding Norovirus polypeptides
The antibodies comprise a heavy chain variable domain with defined CDR1, CDR2, and CDR3 sequences from SEQ ID NOs: 1-30 that specifically bind NoV polypeptides, particularly VP1 from Genogroup I or II Noroviruses such as Norwalk and MD2004 strains.
Method for detecting Norovirus in a sample using the VHH antibodies
A diagnostic method involving contacting a biological or environmental sample with the isolated VHH monoclonal antibody or antigen binding fragments that specifically bind Norovirus, then detecting the binding to indicate presence of the virus.
Compositions comprising therapeutic VHH monoclonal antibodies
Pharmaceutical compositions comprising a therapeutically effective amount of the isolated VHH monoclonal antibodies with defined heavy chain variable domains that specifically bind Norovirus polypeptides, formulated for routes including enteric administration and optionally combined with antiviral agents.
Kits for detecting Norovirus comprising specified VHH monoclonal antibodies
Kits comprising containers with isolated VHH monoclonal antibodies or antigen binding fragments with specified CDR sequences capable of specifically binding Norovirus polypeptides, optionally including second antibodies for detection and components for assays.
The claims define isolated single-domain VHH monoclonal antibodies against Norovirus with specific sequence-defined binding regions, methods for their diagnostic use in detecting Norovirus, therapeutic compositions containing the antibodies, and diagnostic kits including these VHH antibodies, collectively covering detection and treatment of Norovirus infections.
Stated Advantages
The VHH monoclonal antibodies possess high binding affinity and specificity towards Norovirus polypeptides, including VP1, allowing precise detection and neutralization.
Due to their small size and stability, VHH antibodies can bind to epitopes inaccessible to conventional antibodies, enhancing diagnostic and therapeutic potential.
The antibodies can be produced recombinantly in various expression systems, including prokaryotic and eukaryotic hosts, allowing scalable production.
The disclosed antibodies can neutralize multiple Norovirus genogroups and variants, facilitating broad spectrum prophylaxis or therapy.
The diagnostic methods employing these antibodies allow sensitive and specific detection of Norovirus in biological and environmental samples.
Documented Applications
Detection of Norovirus presence in biological samples such as stool, blood, serum, plasma, sputum, nasopharyngeal secretions, spinal fluid, or urine, and environmental samples, using the VHH monoclonal antibodies.
Use of the VHH antibodies in diagnostic assays for Norovirus, including to detect infection or to test vaccine efficacy by confirming immunogenic antigen presence.
Therapeutic treatment of Norovirus infections in subjects at risk or infected, including immunocompromised patients, children, and elderly, by administering therapeutically effective amounts of VHH monoclonal antibodies or nucleic acids encoding them.
Formulations of the VHH antibodies for systemic, parenteral, intramuscular, or enteric administration for prophylaxis or treatment of Norovirus infections.
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