Methods for the production of therapeutic, diagnostic, or research antibodies
Inventors
Assignees
Publication Number
US-12065672-B2
Publication Date
2024-08-20
Expiration Date
2038-08-30
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Abstract
Down-regulating autoimmune regulator (AIRE) function in B cells to produce antibodies is described. The antibodies can be class-switched, high affinity, and neutralizing, and have a high degree of somatic hypermutations, even in the framework region, as compared to antibodies produced in the absence of AIRE downregulation.
Core Innovation
The patent discloses methods and systems for improving the generation of antibodies by down-regulating the function of the autoimmune regulator (AIRE) in B cells. The core innovation is that B cells with reduced AIRE function, when exposed to specific antigens, exhibit increased rates of somatic hypermutation (SHM) and class switch recombination (CSR), leading to the production of high affinity, class-switched, and neutralizing antibodies. These antibodies possess a higher degree of somatic hypermutations, including in both complementarity-determining regions (CDRs) and framework (FR) regions, compared to antibodies produced without AIRE downregulation.
The background emphasizes that a diversified antibody repertoire is essential for effective immunity and prevention of autoimmunity, with B cells undergoing affinity maturation through SHM and CSR in germinal centers. However, the process is regulated in part by AIRE, which acts as a checkpoint, limiting excessive antibody diversification. The problem addressed by the invention is to overcome this checkpoint to generate enhanced antibodies for therapeutic, diagnostic, or research purposes, particularly when a heightened or specific immune response is desirable.
The system allows for B cell modification via gene-editing methods, such as CRISPR-Cas, TALENs, MegaTALs, or zinc finger nucleases, to inhibit AIRE expression or function. By applying these methods in vitro or in vivo, and by exposing the modified B cells to antigens—with optional adjuvants—the process results in the development of antibodies with improved functional characteristics. The methods are applicable to the primary and recall immune response, enabling the generation of monoclonal, polyclonal, human, non-human, or humanized antibodies with enhanced SHM and CSR, for use in multiple biomedical contexts.
Claims Coverage
There is one independent claim detailing the inventive features related to methods for increasing somatic hypermutation and class switch recombination during antibody production.
Method for increasing somatic hypermutation (SHM) and class switch recombination (CSR) during antibody production by using B cells with down-regulated AIRE function
This feature involves: - Selecting a population of B cells with down-regulated autoimmune regulator (AIRE) function that has been contacted with an antigen. - Stimulating the B cell population within in vitro culture conditions, wherein stimulation includes adding one or more of CD40L, IL-4, IFN-γ, or TGF-α. - The increase in SHM and CSR occurs in relation to antibodies produced by a population of B cells with normal AIRE function contacted with the same antigen under comparable conditions.
The claim centers on producing antibodies with enhanced maturation and diversity by specifically manipulating AIRE function in B cells and providing defined conditions for stimulation, distinguishing the method from standard approaches that do not involve AIRE downregulation.
Stated Advantages
Produces antibodies with increased somatic hypermutation (SHM) and class switch recombination (CSR), resulting in higher affinity and/or neutralizing antibodies.
Allows for the development of antibodies with increased mutations in both the CDR and FR regions compared to methods not using AIRE downregulation.
Facilitates the generation of antibodies for clinical and biomedical research applications, including therapeutic, diagnostic, and research uses.
Documented Applications
Therapeutic uses, including antibody-based therapeutics for cancer, autoimmune, and inflammatory diseases.
Diagnostic and research applications, such as flow cytometry, imaging, immunohistochemistry, and western blot.
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