Single-domain VHH 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-10000556-B2
Publication Date
2018-06-19
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 relates to isolated VHH monoclonal antibodies that specifically bind to a Norovirus polypeptide, including Genogroup I and Genogroup II NoVs, such as Norwalk virus (NV) and MD2004 virus. The antibodies can specifically bind VP1, including its P1 or P2 subdomains, and can be neutralizing. The disclosure also covers nucleic acids encoding these antibodies, expression vectors, isolated host cells expressing the nucleic acids, compositions including the antibodies or nucleic acids, and methods for detecting norovirus presence or infection and for treating or preventing NoV infection.
The problem being solved is the need for reagents capable of detecting and treating NoV infections. Norovirus is a leading cause of epidemic gastroenteritis and poses a significant health burden, especially in vulnerable populations such as young children, the elderly, and immunocompromised individuals. NoV infections lack specific therapeutic options and vaccines, posing challenges in diagnosis and treatment. Moreover, the absence of an in vitro culture system limits antigenic studies and vaccine development. Thus, the invention provides new recombinant single-domain antibodies from llamas that can be used for detection, immunoprophylaxis, and treatment of NoV infections.
Claims Coverage
The claims include multiple independent claims directed to isolated VHH monoclonal antibodies specific for Norovirus polypeptides, nucleic acids encoding these antibodies, compositions containing them, and methods of detection and treatment of NoV infection using these antibodies.
VHH monoclonal antibodies with specific heavy chain CDR sequences
Isolated VHH monoclonal antibodies comprising heavy chain variable domains with defined complementarity determining regions (CDR1, CDR2, and CDR3) as specified by amino acid positions in SEQ ID NOs 1-21 or 22-30, which specifically bind Norovirus polypeptides.
Antibody forms and labeling
The antibodies can be IgG, humanized or chimeric monoclonal antibodies. The antibodies or antigen binding fragments may be labeled with detectable moieties such as fluorescent, enzymatic, or radioactive labels.
Compositions including antibodies and nucleic acids
Pharmaceutical compositions comprising effective amounts of the antibodies or nucleic acids encoding them along with pharmaceutically acceptable carriers.
Nucleic acids encoding VHH antibodies and expression vectors
Isolated nucleic acid molecules encoding the monoclonal antibodies, operably linked to promoters, incorporated into expression vectors including viral vectors, and transformed host cells expressing these nucleic acids.
Methods for detecting Norovirus infection
Methods of detecting the presence of Norovirus infection in a biological sample by contacting the sample with an isolated monoclonal antibody or antigen binding fragment and detecting antibody binding.
Methods for inhibiting Norovirus infection
Methods for inhibiting Norovirus infection in subjects by administering a therapeutically effective amount of the isolated antibody, antigen binding fragment, or nucleic acid encoding these molecules, optionally in combination with an anti-viral agent, with optional measuring of viral titer.
The claims cover isolated VHH monoclonal antibodies with defined heavy chain CDR sequences that specifically bind Norovirus polypeptides, compositions and nucleic acids encoding them, expression systems, and methods for detecting and treating Norovirus infections using these antibodies or their encoding nucleic acids.
Stated Advantages
The VHH monoclonal antibodies have high binding affinity and specific binding to Norovirus polypeptides including variants from different genogroups and genotypes.
The small size of VHH antibodies allows them to bind antigenic sites inaccessible to larger antibodies, increasing their utility in detection and treatment.
They are thermostable, stable to extreme pH and proteolytic digestion, poorly antigenic, and can cross biological barriers, such as the blood-brain barrier.
VHH antibodies can be efficiently produced recombinantly in various expression systems.
The disclosed antibodies demonstrate neutralizing activity and blockade of virus binding to host carbohydrates, indicating potential therapeutic application.
Documented Applications
Detection of Norovirus in biological samples to diagnose infection using the antibodies or antigen binding fragments.
Treatment and prevention of Norovirus infection in subjects by administering therapeutically effective amounts of antibodies or nucleic acids encoding the antibodies.
Use of the antibodies in surrogate neutralization assays and vaccine testing to evaluate induction of neutralizing antibodies.
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