Use of vitellogenin for defining and testing novel immunogens in insects
Inventors
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12145965-B2
Publication Date
2024-11-19
Expiration Date
Abstract
The present invention relates to methods of isolating a honey bee antigen that could be useful as a vaccine. The present invention further relates to proteomic methods of identifying antigenic proteins.
Core Innovation
The invention provides a method to isolate a honey bee antigen by using recombinant vitellogenin (rVg) as an affinity capture agent. rVg is formed on a solid support and then contacted with a solubilized inactivated honey bee pathogen so that antigenic proteins or fragments bind to the rVg. The bound components are recovered by eluting the inactivated honey bee pathogen to isolate the honey bee antigen.
The disclosed approach is grounded in isolating antigenic components from inactivated honey bee pathogens, including American foulbrood (AFB)-causing Paenibacillus larvae, and optionally other honey bee pathogens described in the document. The bound proteins can then be identified using analytical techniques such as electrophoresis and Western blot, and further protein identification using standard protein analytical methods.
The disclosure further proposes workflows for antigen and immunogen discovery in which antibodies are replaced by labeled rVg in high-throughput screening, including phage display or genome screening, to identify proteins or fragments that bind rVg. After identification, the document describes downstream vaccine or bacterin production based on the identified proteins.
Claims Coverage
The independent claim describes a method for affinity-based isolation of a honey bee antigen using immobilized recombinant vitellogenin and elution of bound inactivated honey bee pathogen material. The inventive features are centered on immobilized rVg, contacting with solubilized inactivated honey bee pathogen, and eluting to isolate the honey bee antigen, with dependent refinements adding support formats, pathogen solubilization/inactivation, and identity determination features.
Affinity capture using immobilized recombinant vitellogenin
Forming recombinant vitellogenin (rVg) on a solid support and contacting the rVg-bound solid support with a solubilized inactivated honey bee pathogen to enable binding of antigenic material.
Elution of bound inactivated honey bee pathogen to isolate honey bee antigen
Eluting the inactivated honey bee pathogen to isolate the honey bee antigen.
Recombinant vitellogenin defined by amino acid sequence
Using an rVg protein having the amino acid sequence of SEQ ID NO: 1.
Selected solid support formats
Using a solid support selected from a chromatographic support, a sepharose CL column, or an ion exchange column.
Pathogen solubilization and inactivation
Further inactivating honey bee pathogens by sonicating or solubilizing them using non-ionic detergents.
Honey bee pathogen selection from specified Paenibacillus species
Selecting the honey bee pathogen from one or more specified Paenibacillus species including Paenibacillus larvae and other enumerated Paenibacillus species.
Identity determination using protein analytical techniques
Determining the identity of the isolated inactivated honey bee pathogen using electrophoresis, high performance liquid chromatography, mass spectrometry, and/or protein sequencing.
The claim coverage focuses on immobilized rVg for affinity capture of solubilized inactivated honey bee pathogen material, elution to isolate honey bee antigen, and optional identity determination using protein analytical techniques.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Affinity capture and isolation of honey bee antigens from solubilized inactivated honey bee pathogens, including American foulbrood (AFB)-causing Paenibacillus larvae.
Antigen and immunogen discovery workflows using labeled rVg in high-throughput screening, including phage display or genome screening.
Vaccine or bacterin production based on identified proteins.
Insect feed delivery of the vaccine or bacterin to queen bee wafers, sugar feed, or artificial diet, with optional inclusion of TLR-binding CpGs.
Interested in licensing this patent?