Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fc Variants Having Decreased Affinity for FcyRIIc

a variant and affinity technology, applied in the field of new optimized fc variants, can solve the problems of unsatisfactory potency of antibodies as anti-cancer agents, another level of complexity, and biotherapeutics consume essentially all of the available manufacturing capacity, so as to improve the function and/or solution properties, improve the affinity, and facilitate the effect of effector function

Inactive Publication Date: 2007-10-11
XENCOR INC
View PDF98 Cites 91 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides Fc variants that are optimized for therapeutic purposes. These Fc variants have been designed to have improved properties such as increased effector function, enhanced antibody-dependent cellular cytotoxicity, and improved solubility. The invention also provides Fc variants that have been characterized at various positions in the Fc region of the antibody. These Fc variants can be used in therapeutic proteins such as antibodies or Fc fusion proteins. Overall, the invention provides a novel way to improve the function of Fc variants in antibodies and related proteins.

Problems solved by technology

Yet another level of complexity is the existence of a number of FcγR polymorphisms in the human proteome.
Despite this arsenal of anti-tumor weapons, the potency of antibodies as anti-cancer agents is unsatisfactory, particularly given their high cost.
Antibodies must be expressed in mammalian cells, and the currently marketed antibodies together with other high-demand biotherapeutics consume essentially all of the available manufacturing capacity.
First, it dramatically raises the cost of goods to the producer, a cost that is passed on to the patient.
Second, it hinders industrial production of approved antibody products, limiting availability of high demand therapeutics to patients.
Finally, because clinical trials require large amounts of a protein that is not yet profitable, the insufficient supply impedes progress of the growing antibody pipeline to market.
This may result in reduced or even lack of effector function because, as discussed above, the carbohydrate structure can significantly impact FcγR and complement binding.
A potentially greater problem with nonhuman glycoforms may be immunogenicity; carbohydrates are a key source of antigenicity for the immune system, and the presence of nonhuman glycoforms has a significant chance of eliciting antibodies that neutralize the therapeutic, or worse cause adverse immune reactions.
Thus the efficacy and safety of antibodies produced by transgenic plants and animals remains uncertain.
For complex proteins such as antibodies there are a number of obstacles to bacterial expression, including folding and assembly of these complex molecules, proper disulfide formation, and solubility, stability, and functionality in the absence of glycosylation because proteins expressed in bacteria are not glycosylated.
However the ultimate utility of bacterially expressed antibodies as therapeutics remains hindered by the lack of glycosylation, which results in lack effector function and may result in poor stability and solubility.
This will likely be more problematic for formulation at the high concentrations for the prolonged periods demanded by clinical use.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fc Variants Having Decreased Affinity for FcyRIIc
  • Fc Variants Having Decreased Affinity for FcyRIIc
  • Fc Variants Having Decreased Affinity for FcyRIIc

Examples

Experimental program
Comparison scheme
Effect test

example 1

Molecular Biology and Protein Expression / Purification

[0250] The majority of experimentation on the Fc variants was carried out in the context of the anti-cancer antibody alemtuzumab (Campath®, a registered trademark of Ilex Pharmaceuticals LP). Alemtuzumab binds a short linear epitope within its target antigen CD52 (Hale et al., 1990, Tissue Antigens 35:118-127; Hale, 1995, Immunotechnology 1:175-187). Alemtuzumab has been chosen as the primary engineering template because its efficacy is due in part to its ability to recruit effector cells (Dyer et al., 1989, Blood 73:1431-1439; Friend et al., 1991, Transplant Proc 23:2253-2254; Hale et al., 1998, Blood 92:4581-4590; Glennie et al., 2000, Immunol Today 21:403-410), and because production and use of its antigen in binding assays are relatively straightforward. In order to evaluate the optimized Fc variants of the present invention in the context of other antibodies, select Fc variants were evaluated in the anti-Her2 antibody trastu...

example 2

Fc Ligand Binding Assays

[0254] Binding to the human Fc ligands FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, C1q, and FcRn was measured for the designed Fc variants. Binding affinities were measured using an AlphaScreen™ assay (Amplified Luminescent Proximity Homogeneous Assay (ALPHA), PerkinElmer, Wellesley, MA), a bead-based luminescent proximity assay. Laser excitation of a donor bead excites oxygen, which if sufficiently close to the acceptor bead generates a cascade of chemiluminescent events, ultimately leading to fluorescence emission at 520-620 nm. WT alemtuzumab antibody was biotinylated by standard methods for attachment to streptavidin donor beads, and GST-tagged FcγRs and FcRn were bound to glutathione chelate acceptor beads. For the C1q binding assay, untagged C1q protein was conjugated with Digoxygenin (DIG, Roche) using N-hydrosuccinimide (NHS) chemistry and bound to DIG acceptor beads. For the protein A binding assay, protein A acceptor beads were purchased directly f...

example 3

ADCC of Fc Variants

[0267] In order to determine the effect on effector function, cell-based ADCC assays were performed on select Fc variants. ADCC was measured using the DELFIA® EuTDA-based cytotoxicity assay (Perkin Elmer, MA) with purified human peripheral blood monocytes (PBMCs) as effector cells. Target cells were loaded with BATDA at 1×106 cells / ml, washed 4 times and seeded into 96-well plate at 10,000 cells / well. The target cells were then opsonized using Fc variant or WT antibodies at the indicated final concentration. Human PBMCs, isolated from buffy-coat were added at the indicated fold-excess of target cells and the plate was incubated at 37° C. for 4 hrs. The co-cultured cells were centrifuged at 500×g, supernatants were transferred to a separate plate and incubated with Eu solution, and relative fluorescence units were measured using a Packard Fusion™α-FP HT reader (Packard Biosciences, IL). Samples were run in triplicate to provide error estimates (n=3, ±S.D.). PBMCs ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
timeaaaaaaaaaa
timeaaaaaaaaaa
Login to View More

Abstract

The present invention relates to Fc variants having decreased affinity for FcγRIIc, methods for their generation, Fc polypeptides comprising optimized Fc variants, and methods for using optimized Fc variants.

Description

[0001] This application is a continuation of U.S. application Ser. No. 11 / 124,620, filed May 5, 2005, which claims benefit under 35 U.S.C. §119(e) to U.S. Ser. No. 60 / 568,440, filed May 5, 2004; Ser. No. 60 / 589,906 filed Jul. 20, 2004; Ser. No. 60 / 627,026 filed Nov. 9, 2004; Ser. No. 60 / 626,991 filed Nov. 10, 2004; Ser. No. 60 / 627,774 filed Nov. 12, 2004, Ser. No. 60 / 531,752, filed Dec. 22, 2003; and, Ser. No. 60 / 531,891, filed Dec. 22, 2003; and which is continuation-in-part of U.S. Ser. No. 10 / 822,231, filed Mar. 26, 2004; which is continuation-in-part of Ser. No. 10 / 672,280, filed Sep. 26, 2003; which is a continuation-in-part of Ser. No. 10 / 379,392, filed Mar. 3, 2003, all of which are incorporated by reference in their entirety.FIELD OF THE INVENTION [0002] The present invention relates to novel optimized Fc variants, engineering methods for their generation, and their application, particularly for therapeutic purposes. BACKGROUND OF THE INVENTION [0003] Antibodies are immunolo...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/16C07K16/00
CPCA61K2039/505C07K2317/34C07K16/2887C07K16/2893C07K16/32C07K2317/24C07K2317/41C07K2317/71C07K2317/72C07K2317/732C07K2317/734C07K2317/77C07K2317/52C07K16/18C07K16/2863
Inventor LAZAR, GREGORY ALANDANG, WEIDESJARLAIS, JOHN R.KARKI, SHER BAHADURVAFA, OMIDHAYES, ROBERT
Owner XENCOR INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com