Identification of antibodies specific for lyssaviruses and methods of their use
Inventors
Assignees
Centers of Disease Control and Prevention CDC • US Department of Health and Human Services
Publication Number
US-9115187-B2
Publication Date
2015-08-25
Expiration Date
2031-10-18
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Abstract
Described herein is a method of identifying a monoclonal antibody (or antigen-binding fragment thereof) that specifically binds a plurality of lyssaviruses for use in post-exposure rabies prophylaxis or in the treatment of clinical rabies. The method includes using a naïve antibody phage display library to screen for phage clones that bind whole recombinant rabies virus or cells expressing glycoprotein from multiple lyssaviruses (such as RABV, MOKV and WCBV) and/or specifically bind recombinant glycoprotein from different lyssaviruses.
Core Innovation
The invention relates to a method of identifying monoclonal antibodies or antigen-binding fragments thereof that specifically bind a plurality of lyssaviruses. The method uses a naïve antibody phage display library screened against multiple lyssavirus glycoproteins or viruses expressing these glycoproteins. Identified antibodies can bind to at least two different lyssaviruses, such as rabies virus (RABV), Mokola virus (MOKV), and West Caucasian bat virus (WCBV), among others. These antibodies are useful for post-exposure rabies prophylaxis or treatment of clinical rabies.
The background problem addressed is that rabies remains a fatal but preventable disease with a significant global burden. Current post-exposure prophylaxis requires administration of immunoglobulins (RIG) and vaccine, but RIG availability is limited, especially in developing countries, and existing RIG may have side effects or risk of pathogen transmission. Furthermore, existing human monoclonal antibodies often target only genotype 1 lyssaviruses and have limited cross-reactivity to other genotypes. The invention overcomes the limitation of using immunized humans for antibody libraries which biases antibody diversity primarily to genotype 1 rabies virus, by instead utilizing naïve antibody libraries to identify broadly neutralizing antibodies.
The invention details screening strategies involving panning naïve human VH domain phage display libraries against viruses or cells expressing multiple lyssavirus glycoproteins, allowing selection of antibodies broadly reactive across multiple lyssavirus genotypes. Monoclonal antibodies or fragments identified can be cloned into IgG expression vectors for production and used for therapeutics. Recombinant viruses expressing multiple glycoproteins and cell lines expressing different lyssavirus glycoproteins facilitate this identification process.
Claims Coverage
The claims include one independent method claim with several related dependent claims. The key inventive features cover a method of identifying monoclonal antibodies or fragments that bind multiple lyssavirus species by sequential screening of a naïve antibody phage display library on cell lines expressing different lyssavirus glycoproteins.
Sequential screening of a naïve antibody phage display library on cell lines expressing different lyssavirus glycoproteins
A method of identifying a monoclonal antibody or antigen-binding fragment that specifically binds at least two different lyssavirus species by screening a naïve antibody phage display library sequentially on at least two different cell lines, each expressing a different lyssavirus glycoprotein selected from RABV, MOKV, WCBV, LBV, and DUVV glycoproteins.
Use of a naïve human VH domain library
The phage display library used in the method is a naïve human variable heavy (VH) domain library for antibody selection.
Antigen-binding fragment comprising a VH domain
The identified antigen-binding fragment includes a VH domain that binds to at least two lyssavirus species.
Cloning into an IgG expression vector
The antigen-binding fragment comprising the VH domain is cloned into an IgG expression vector, which can also include a variable light (VL) domain from a rabies virus-specific antibody.
Selection using ELISA
The method includes selecting phage display clones based on specific binding detected by ELISA to lyssavirus glycoprotein, whole virus, or both.
Screening for lyssavirus neutralization
The method further includes screening the selected phage display clones for their ability to neutralize lyssavirus infectivity.
The claims cover a comprehensive method of identifying lyssavirus cross-reactive monoclonal antibodies using sequential panning of naïve human VH domain libraries on cell lines expressing different lyssavirus glycoproteins, with selection and validation steps including ELISA and neutralization assays, as well as cloning for expression.
Stated Advantages
Overcomes limitations of current RIG by providing monoclonal antibodies that neutralize multiple lyssavirus species.
Antibodies can be produced in a cost-effective manner with reduced risk of immune response or pathogen transmission.
Improves breadth of virus neutralization, including lyssaviruses outside the typical vaccine cross-protection group.
Facilitates development of broadly neutralizing human monoclonal antibodies derived from naïve libraries, providing improved prophylaxis and clinical treatment options.
Documented Applications
Use of monoclonal antibodies for post-exposure rabies prophylaxis.
Treatment of clinical rabies in subjects exposed to or infected by lyssaviruses.
Administration of identified antibodies or antibody fragments systemically or locally (e.g., to wound sites) to inhibit rabies virus replication or spread.
Use of antibody compositions in conjunction with rabies vaccines.
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