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Immunomodulatory proteins with tunable affinities

A technology of immunomodulatory proteins and immunoglobulins, which can be used in peptide/protein components, animal/human proteins, medical preparations containing active ingredients, etc., and can solve problems such as lack of important attributes

Inactive Publication Date: 2018-04-27
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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  • Immunomodulatory proteins with tunable affinities
  • Immunomodulatory proteins with tunable affinities
  • Immunomodulatory proteins with tunable affinities

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0279] The implementation methods provided include:

[0280] Embodiment 1. In some embodiments, an immunomodulatory protein is provided, which contains at least one non-immunoglobulin affinity modified immunoglobulin superfamily (IgSF) domain, the immunoglobulin superfamily domain containing a wild-type IgSF domain One or more amino acid substitutions in wherein: at least one affinity-modified IgSF domain has enhanced binding to at least two associated binding partners compared to the wild-type IgSF domain; and at least one affinity-modified IgSF domain is specific To non-competitively bind at least two related binding partners.

[0281] Embodiment 2. In some other embodiments of embodiment 1, at least two cognate binding partners are cell surface molecules expressed on the surface of mammalian cells.

[0282] Embodiment 3. In some other embodiments of Embodiment 2, the cell surface molecule is expressed in a cis configuration or a trans configuration.

[0283] Embodiment 4. In s...

Embodiment 1

[0477] Generation of mutant DNA constructs of IgSF domain

[0478] Example 1 describes the generation of mutant DNA constructs of human CD80, CD86, ICOSL and NKp30 IgSF domains for translation and expression on the surface of yeast as a yeast display library.

[0479] A. Degenerate library

[0480] For libraries that are fully or partially randomized with degenerate codons for specific residues of the target protein, human CD80 (SEQ ID NO: 28), ICOSL (SEQ ID NO: 32) and NKp30 (SEQ ID NO: 54) cells A collection of overlapping oligonucleotides with a length of more than 80 base pairs (bp) from the coding DNA of the outer domain (ECD) was purchased from Integrated DNA Technologies (Clover, Iowa, USA). In order to generate a library of multiple variants of each ECD, the oligonucleotide contains the required degenerate codons at the required amino acid positions. Degenerate codons are generated using the algorithm located at URL: rosettadesign.med.unc.edu / SwiftLib / .

[0481] Generally, t...

Embodiment 2

[0488] DNA library introduced into yeast

[0489] Example 2 describes the introduction of CD80, CD86, ICOSL, and NKp30 DNA libraries into yeast.

[0490] In order to introduce degenerate and random library DNA into yeast, electroporation competent cells of yeast strain BJ5464 (ATCC.org; ATCC No. 208288) were prepared, and they were basically as described on Gene Pulser II (BioRad, USA). The electroporation is performed using the electroporation ready-to-use DNA from the above steps (Colby, DW, etc., 2004 Methods Enzymology 388, 348-358). The only exception is that the transformed cells were grown in non-inducible minimal selective SCD-Leu medium to accommodate the LEU2 selectable marker carried by the modified plasmid pBYDS03.

[0491] The library size was determined by plating a dilution of freshly recovered cells on SCD-Leu agar plates, and then inferring the library size from the number of individual colonies from the plating that produced at least 50 colonies per plate. The rem...

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Abstract

Provided are immunomodulatory proteins and nucleic acids encoding such proteins. The immunomodulatory proteins 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 requires U.S. Provisional Application No. 62 / 149,437, filed on April 17, 2015, entitled "Immune Regulatory Proteins with Adjustable Affinity", and filed on September 14, 2015, entitled "Immune Regulatory Proteins with Adjustable Affinity" The priority of "Immune Regulatory Proteins" of US Provisional Application No. 62 / 218,534, the contents of which are incorporated herein by reference in their entirety. [0003] Included in sequence listing by reference [0004] This application is submitted with the sequence table in electronic format. The provided sequence listing name is 761612000140SEQLIST.TXT, created on April 15, 2016, and its file size is 377kb. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety. Technical field [0005] The present invention relates to therapeutic proteins used in the treatment of cancer and immune diseases to adjust im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63A61K38/17A61K35/00
CPCC07K14/70532C07K2319/30C07K14/70503C07K14/70521C07K14/70596A61P1/04A61P11/00A61P11/06A61P13/10A61P17/00A61P17/06A61P19/02A61P25/00A61P29/00A61P35/00A61P35/02A61P37/02A61P37/04A61P37/06A61P7/00Y02A50/30A61K38/00C07K14/715C07K2319/73G01N33/564G01N33/57484
Inventor R·斯旺森M·科纳克
Owner ALPINE IMMUNE SCI INC
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