Norovirus antibodies
Inventors
HANSMAN, Grant • KOROMYSLOVA, Anna
Assignees
Publication Number
US-10233232-B2
Publication Date
2019-03-19
Expiration Date
2035-10-14
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
The present invention relates to a binding polypeptide specifically binding to the amino acid sequence W-V-N-X1-F-Y-X2 (SEQ ID NO: 1), wherein X1 represents any amino acid, preferably Q or P, and wherein X2 represents any amino acid, preferably, T or S in a norovirus polypeptide. The present invention further relates to polynucleotide encoding a binding polypeptide of the present invention an to a host cell comprising the same or the polynucleotide of the invention. The present invention further relates to a method of detecting the presence of a norovirus capsid polypeptide in a sample and to kits, devices, and uses making use of the binding peptide of the invention.
Core Innovation
The invention is directed to binding polypeptides that specifically recognize the amino acid sequence W-V-N-X1-F-Y-X2 (SEQ ID NO:1) present in norovirus polypeptides, where X1 can be any amino acid but is preferably Q or P, and X2 can be any amino acid but is preferably T or S. The primary focus is on binding polypeptides that can selectively attach to this conserved sequence region found especially in norovirus capsid polypeptides. Such binding polypeptides include antibodies, particularly single-domain antibodies (VHH or nanobodies) that can specifically bind this sequence with high affinity.
The invention also addresses polynucleotides encoding such binding polypeptides, host cells containing these polynucleotides or polypeptides, and methods employing these binding polypeptides for diagnostic, therapeutic, and preventive applications. The methods include detecting norovirus capsid proteins in samples and potentially inactivating norovirus particles by binding them, thus preventing infection or spread. Kits and devices incorporating these binding polypeptides for norovirus detection and treatment are also part of the invention.
The problem being solved by the invention arises from the difficulty in broadly detecting and treating human noroviruses due to their genetic and antigenic diversity and the inability to cultivate them effectively in cell culture, which hampers vaccine and antiviral development. Prior antibodies either lacked broad reactivity or could not effectively prevent infection. There remained a need for improved antibodies capable of broad recognition across norovirus genotypes, facilitating better diagnostic tools and potential therapeutic interventions. The invention seeks to provide such improved antibodies that overcome these limitations.
Claims Coverage
The patent contains 10 inventive features extracted from the independent claims focused on a single-domain antibody binding specifically to a defined norovirus amino acid sequence, corresponding polynucleotides, host cells, and kits or compositions using the antibody.
Single-domain antibody specifically binding to defined norovirus sequence
A single-domain antibody (VHH) that binds specifically to the amino acid sequence W-V-N-X1-F-Y-X2 (SEQ ID NO:1), where X1 is preferably Q or P and X2 is preferably T or S, comprising complementarity determining regions (CDRs): CDR1 of GSIFSIYA or GSIFSIYL, CDR2 of ISSGGGTN, and CDR3 of KREDYSAYAPPSGS or KREDFSAYAPPSGS.
Specific binding to extended norovirus polypeptide sequences
The single-domain antibody binds specifically to norovirus sequences selected from extended motifs including F-X6-X5-X4-X3-W-V-N-X1-F-Y-X2 or longer contiguous sequences encompassing SEQ ID NO:1.
Single-domain antibody encoded by nucleic acid with at least 70% sequence identity to SEQ ID NO:5
The VHH is encoded by a nucleotide sequence having at least 70% sequence identity to SEQ ID NO:5, enabling production of the antibody using nucleic acid molecules with significant homology.
Single-domain antibody comprising specific set of CDRs
The antibody comprises the complementarity determining regions CDR1: GSIFSIYA, CDR2: ISSGGGTN, and CDR3: KREDYSAYAPPSGS, conferring specificity to the norovirus sequence.
Single-domain antibody comprising the amino acid sequence of SEQ ID NO:9
The antibody comprises the amino acid sequence as set forth in SEQ ID NO:9, providing a defined structure for binding to the target norovirus epitope.
Norovirus polypeptide targeted is a capsid polypeptide
The norovirus polypeptide to which the antibody binds is a capsid polypeptide, emphasizing the functional targeting of the virus’s structural component.
Host cell comprising the single-domain antibody
A host cell that includes the single-domain antibody, enabling recombinant expression and production of the antibody.
Kit for diagnosing, preventing, or treating norovirus infection comprising the single-domain antibody
A kit containing the single-domain antibody for use in diagnosing, preventing, or treating norovirus infections.
Single-domain antibody with at least 80% sequence identity to SEQ ID NO:9
The antibody comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:9, allowing for conservative sequence variations while retaining functionality.
Single-domain antibody comprising amino acid substitutions A33L, Y100F, or both
The antibody comprising the sequence of SEQ ID NO:9 with one or more amino acid substitutions selected from A33L, Y100F, and the double mutation A33L/Y100F to enhance binding properties.
The claims principally cover a single-domain antibody targeting a conserved norovirus capsid amino acid sequence with defined CDRs, corresponding polynucleotides, variants with sequence identity and specific mutations, host cells for antibody expression, and diagnostic and therapeutic kit compositions incorporating this antibody, providing broad coverage of antibody production, sequence characteristics, and use.
Stated Advantages
The binding polypeptide has the capacity to broadly recognize diverse genogroup II norovirus members and some genogroup I members, facilitating broad diagnostic detection.
The antibody potentially inhibits norovirus infection regardless of genotype, offering therapeutic potential.
The antibody can inactivate norovirus particles by binding to a specific site, preventing further spread of infection.
Nanobodies are easy and cost-effective to produce, enabling low-cost norovirus detection kits.
The small size of nanobodies allows for high-density application on solid surfaces, improving assay sensitivity and suitability for techniques like immunochromatography.
Documented Applications
Use in diagnostic tests to detect norovirus capsid polypeptides in various samples, including stool, blood, plasma, urine, saliva, and other body fluids.
Therapeutic use for treating or preventing norovirus infections in subjects via administration of the binding polypeptide.
Use for inactivating norovirus particles on objects or in subjects to prevent infection or spread.
Incorporation into kits and devices for diagnosing, preventing, or treating norovirus infections.
Potential use for sanitization and disinfection purposes by applying the binding polypeptide to objects suspected to harbor norovirus.
Interested in licensing this patent?