Invented by Frank Kowalewsky, Mirko Ritter, Kay-Gunnar Stubenrauch, Uwe Wessels, Hoffmann La Roche Inc

The market for Anti-variant Fc region antibodies and methods of use Introduction: Antibodies play a crucial role in the field of biotechnology and medicine. They are widely used as therapeutic agents, diagnostic tools, and research reagents. In recent years, there has been a growing interest in developing antibodies that specifically target the Fc region of immunoglobulins, known as anti-variant Fc region antibodies. These antibodies have shown great potential in various applications, including cancer therapy, autoimmune diseases, and immunotherapy. This article will explore the market for anti-variant Fc region antibodies and discuss their methods of use. Market Overview: The market for anti-variant Fc region antibodies is witnessing significant growth due to the increasing demand for targeted therapies and personalized medicine. The global antibody therapeutics market is expected to reach $300 billion by 2025, with a compound annual growth rate (CAGR) of 14.5%. Anti-variant Fc region antibodies are expected to contribute significantly to this growth, driven by their unique ability to modulate immune responses and enhance therapeutic efficacy. Applications in Cancer Therapy: One of the most promising applications of anti-variant Fc region antibodies is in cancer therapy. These antibodies can be engineered to selectively bind to cancer cells, leading to their destruction through various mechanisms such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Additionally, anti-variant Fc region antibodies can be used to enhance the efficacy of existing cancer therapies, such as immune checkpoint inhibitors, by modulating the immune response and overcoming resistance mechanisms. Autoimmune Diseases and Immunotherapy: Anti-variant Fc region antibodies also hold great potential in the treatment of autoimmune diseases. By targeting specific Fc region variants, these antibodies can modulate the immune response and suppress the activity of autoantibodies responsible for autoimmune disorders. This approach offers a more targeted and potentially safer alternative to traditional immunosuppressive therapies. Furthermore, anti-variant Fc region antibodies can be utilized in immunotherapy, particularly in the field of infectious diseases. By targeting specific Fc region variants, these antibodies can enhance the clearance of pathogens and improve immune responses. This has significant implications for the development of novel vaccines and the treatment of viral infections, such as HIV and COVID-19. Methods of Use: Anti-variant Fc region antibodies can be generated through various methods, including hybridoma technology, phage display, and recombinant DNA technology. Hybridoma technology involves fusing antibody-producing B cells with immortalized myeloma cells to create hybridoma cell lines that produce monoclonal antibodies. Phage display utilizes bacteriophages to display antibody fragments, allowing for the selection of specific binders against Fc region variants. Recombinant DNA technology involves the cloning and expression of antibody genes in host cells, enabling the production of large quantities of recombinant antibodies. Once generated, anti-variant Fc region antibodies can be used in various applications. They can be administered as therapeutic agents, either alone or in combination with other drugs, to treat diseases such as cancer and autoimmune disorders. Additionally, these antibodies can be used as research reagents to study the Fc region biology, immune responses, and antibody engineering. Conclusion: The market for anti-variant Fc region antibodies is rapidly expanding, driven by the increasing demand for targeted therapies and personalized medicine. These antibodies have shown great promise in cancer therapy, autoimmune diseases, and immunotherapy. With advancements in antibody engineering and production technologies, the development and commercialization of anti-variant Fc region antibodies are expected to grow significantly in the coming years. This will not only revolutionize the field of biotechnology but also improve patient outcomes and quality of life.

The Hoffmann La Roche Inc invention works as follows

The invention provides antivariant Fc region antibodies that specifically bind an antibody with the mutations P329G, P329G/L234A/L235A, or I253A/H310A/H435A within the Fc region. It also includes methods for using these antibodies.

Background for Anti-variant Fc region antibodies and methods of use

Since the discovery of the first monoclonal antibody by Koehler & Milstein in 1974, a great deal of effort has been put into the development and testing of antibodies that are suitable for human therapy. The first monoclonal antibody that was available in the market had been created by mice and rats. When these antibodies were used to treat humans, they caused unwanted side-effects due to antirodent antibodies. Many efforts have been made to reduce or eliminate such unwanted side effects.

The first human trials will not be possible until a large number of monoclonal humanized antibodies are studied on experimental animals.

Experimental animals are needed to study important criteria such as bioavailability, antibody clearance and many others. In many of these studies, the amount of therapeutic antibodies in relation to the host’s antibodies is quantified. Most often, mammals are used for experiments. Toxicology is often first tested on rodents such as mice or rats. Even monkeys are included in pre-clinical drug studies at the advanced stages of drug research, before the drug is introduced to humans.

The circulation of mammals contains between 10 and 30 milligrams of immunoglobulin.

The levels of monoclonal therapeutic antibodies in serum should range from 1 nanogram to 100 micrograms per ml. It is therefore necessary to detect the therapeutic antibody against a background that contains host antibodies in excess of 100-fold up to 10,000,000-fold. Pharmacologists face a challenging task in detecting a therapeutic human antibody or humanized antibody against a background of host immune globulin. It is also important to note that different therapeutic antibody types may require different assay formats and reagents. The detection of human or humanized antibodies becomes more difficult as the therapeutic antibody gets closer to wild-type human antibody.

The bridging enzyme-linked immunosorbent sandwich test (ELISA), shown in FIG. 1A) is the current state-of-the art format for immunogenicity tests due to its high throughput, sensitivity, and easy application to different projects. (Mikulskis A., and al., J. Immunol. Meth. 365 (2011) 38-49). The reliability of the assay is challenged both by interference caused by oligomeric targets leading to false-positive results (Bautista A. C. et al. Bioanal. 2 (2010) 721-731; Mire-Sluis, A. R., et al., J. Immunol. Meth. 289 (2004) 1-16 (2004); Weeraratne, D. K., et al., J. Immunol. Meth. 396 (2013) 44-55; Zhong, Z. D., et al., J. Immunol. Meth. 355 (2010) 21-28) and the presence in clinical samples of high levels of drug that interferes with labelled drugs and prevents ADAs to generate signals. This leads to false negatives (Mire Sluis A. R. et al. J. Immunol. Meth. 289 (2004) 1-16 (2004); Geng, D., et al., J. Pharm. Biomed. Anal. 39 (2005) 364-375). The detection of ADA in drug immune complexes bound by traditional bridging tests is severely restricted (Mire-Sluis A. R. et al. J. Immunol. Meth. 289 (2004) 1-16 (2004); Geng, D., et al., J. Pharm. Biomed. Anal. 39 (2005) 364-375).

Since bridging tests for the detection of antidrug antibodies are often hampered due to oligomeric target and high drug concentrations it is necessary to improve approaches. Therapeutic antibodies without Fc effector function, e.g. This article reports a drug and target-tolerant immuno complex assay by introducing a Pro329Gly substitution within Fc region, using a capture antigen specific for the substitution, e.g. An anti-PG antibodies and a human soluble Fc receptor are used for detection. receptor for detection. The assay reported here has increased drug and oligomeric targets tolerance compared to conventional bridging (Wessels U., et.al., Bioanalysis 8, (2016) 2135-2145). This method can be used to detect anti-drug antibody even in high drug concentrations because the human Fcgamma soluble receptor, such as the e.g. The human soluble Fc?RI binds specifically to wild-type IgG, but not to Fc-region-modified IgG.

The combination of a bridging test and an ADA Ig subtype assay allows for a detailed ADA characterization. Both assays recognize ADA Ig subtypes differently. The assay described herein complements the conventional bridging test for detailed characterization of individual ADA responses against Fc region-modified therapeutic antibody.

The following steps are included in the assay described here for determining the amount and/or presence of anti-drug antibody(s) in a sample (containing serum).

The following steps are included in the method described here. “One aspect is the determination in vitro of the presence or amount of a specific binding partner that can be bound specifically by a specific binding specificity in a multispecific binder, whereby the binding partner bound to the binder multispecific is depleted before the detection of the binder partner.

The isolated antibody described herein binds specifically to an Fc region containing at positions 253, 301, and 435 the amino acids residue alanine. (numbering is according to Kabat EU Index) It comprises (a) an HVRH1 containing the amino sequence SEQID NO: 09 or 10. (b) an HVRH2 containing the sequence SEQID NO: 12, 13, or 14. (c) an HVRH3 incorporating the sequence SEQID NO: 16, 17, or 18. (a)

This antibody is designated as anti-AAA antibodies in the following.

The following are described in the patent: “One aspect of the invention is an isolated anti-body that specifically binds an Fc region containing at position 329, the amino-acid residue glycine. (and optionally also at positions 234 or 235 the amino-acid residue alanine.) (numbering based on Kabat EU index), comprising: (a) HVR H1 comprising SEQ NO: 20, (b) HVR H2 comprising SEQ NO:

This antibody is designated as anti-PG antibodies in the following.

In one embodiment, the antibody is monoclonal.

In one embodiment, the antibody is either a human antibody, a humanized antibody, or chimeric antibodies.

In one embodiment, the antibody is a fragment of an antibody that binds specifically to the respective Fc-region mutated.

One aspect of the invention is an isolated nucleic acids encoding an antigen as described herein.

One aspect of the invention is that a cell host containing nucleic acids as described herein.

In one embodiment, the host cell is an eukaryotic cellular. In one embodiment, the mammalian eukaryotic cells is used. In one embodiment, the mammalian cells are CHO or HEK cells.

Click here to view the patent on Google Patents.