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Tunable variant immunoglobulin superfamily domains and engineered cell therapy

A technology of immunoglobulin and immunomodulatory protein, applied in the direction of immunoglobulin superfamily, blood/immune system cells, genetically modified cells, etc., can solve problems such as lack of important properties

Inactive Publication Date: 2018-09-07
ALPINE IMMUNE SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both soluble receptors and antibodies lack important properties that are critical for function in IS

Method used

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  • Tunable variant immunoglobulin superfamily domains and engineered cell therapy
  • Tunable variant immunoglobulin superfamily domains and engineered cell therapy
  • Tunable variant immunoglobulin superfamily domains and engineered cell therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0245] The provided implementations include:

[0246] 1. A transmembrane immunomodulatory protein (TIP), comprising:

[0247] (i) an extracellular domain comprising at least one non-immunoglobulin affinity-modified immunoglobulin superfamily (IgSF) domain comprising one or more amino acid substitutions in a wild-type IgSF domain, wherein the at least one affinity modification an IgSF domain that specifically binds at least one cell surface-associated binding partner of said wild-type IgSF domain; and

[0248] (ii) Transmembrane domain.

[0249] 2. The transmembrane immunomodulatory protein of embodiment 1, wherein said at least one cell surface associated binding partner is expressed in a mammalian cell.

[0250] 3. The transmembrane immunomodulatory protein of embodiment 2, wherein the mammalian cell is an antigen presenting cell (APC), a tumor cell or a lymphocyte, which is optionally a T cell.

[0251] 4. The transmembrane immunomodulatory protein according to any one of...

Embodiment 1

[0398] Generation of mutant DNA constructs of the IgSF domain

[0399] Example 1 describes the generation of mutant DNA constructs of human CD80, CD86, ICOSL and NKp30 IgSF domains for translation and expression on the yeast surface as a yeast display library. A. Degenerate library Human CD80 (SEQ ID NO:28), ICOSL (SEQ ID NO:32) and NKp30 (SEQ ID NO:54) cells were fully or partially randomized with degenerate codons for specific residues of the target protein A collection of overlapping oligonucleotides up to 80 base pairs (bp) in length from the coding DNA for the ectodomain (ECD) was ordered from Integrated DNA Technologies (Clarwell, Iowa, USA). To generate a library of multiple variants of each ECD, the oligonucleotides contain the desired degenerate codons at the desired amino acid positions. Degenerate codons were generated using the algorithm at URL: rosettadesign.med.unc.edu / SwiftLib / .

[0400] Typically, the location and degenerate codons of the mutations are selec...

Embodiment 2

[0405] DNA library introduced into yeast

[0406] Example 2 describes the introduction of CD80, CD86, ICOSL, and NKp30 DNA libraries into yeast.

[0407] To introduce degenerate and random library DNA into yeast, electroporation-competent cells of yeast strain BJ5464 (ATCC.org; ATCC No. 208288) were prepared and run on a Gene Pulser II (BioRad, USA) essentially as described. Electroporation was performed using electroporation-ready DNA from the above procedure as described (Colby, D.W. et al., 2004 Methods Enzymology 388, 348-358). The only exception is that transformed cells were grown in non-inducing minimally selective SCD-Leu medium to accommodate the LEU2 selectable marker carried by the modified plasmid pBYDS03.

[0408] Library sizes were determined by plating dilutions of freshly recovered cells on SCD-Leu agar plates and then extrapolating library sizes from the number of individual colonies from platings that produced at least 50 colonies per plate. The remainder o...

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Abstract

Provided are transmembrane immunomodulatory proteins, nucleic acids encoding such proteins and cells engineered with the proteins. The transmembrane immunomodulatory proteins and engineered cells provide therapeutic utility for a variety of immunological and oncological conditions. Compositions and methods for making and using such proteins are provided.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Patent Application No. 62 / 218,531, filed September 14, 2015, entitled "Tunable Variant Immunoglobulin Superfamily Domains and Engineered Cell Therapy," filed April 15, 2016 U.S. Provisional Patent Application No. 62 / 323,608 entitled "ICOS Embryonic Body Variant Immunomodulatory Proteins and Their Uses," filed April 15, 2016, and U.S. Provisional Patent Application entitled "CD80 Variant Immunomodulatory Proteins and Their Uses" Application No. 62 / 323,595, U.S. Provisional Patent Application No. 62 / 367,822, filed July 28, 2016, entitled "CD155 Variant Immunomodulatory Proteins and Uses Thereof," and filed July 28, 2016, entitled "CD112 Variant priority to U.S. Provisional Patent Application No. 62 / 367,819, the respective contents of which are hereby incorporated by reference in their entirety. [0003] Incorporated by reference into the sequence listing [0004] This application...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705
CPCC07K14/70503A61P37/00A61P35/00A61K38/1774A61P1/04A61P11/06A61P17/06A61P19/02A61P25/28A61P29/00A61P37/04Y02A50/30C07K2319/03C07K14/7051A61K39/4631C12N5/0636A61K2239/21A61K39/464496C12N5/0638A61K39/4611C12N5/0635C07K14/70521C12N2510/00C07K14/70517C07K19/00
Inventor R·斯旺森M·科纳克
Owner ALPINE IMMUNE SCI INC
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