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Alk7 binding proteins and uses thereof

A technology for binding proteins and proteins, applied in the field of ALK7 binding proteins and their uses

Active Publication Date: 2019-01-15
ACCELERON PHARMA INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no established cure for type 2 diabetes, but a variety of recognized treatments exist that attempt to delay or lessen the inevitable outcome of the disease

Method used

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  • Alk7 binding proteins and uses thereof
  • Alk7 binding proteins and uses thereof
  • Alk7 binding proteins and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0283] Preparation of ALK7-binding protein

[0284] In some embodiments, the ALK7 binding protein binds the extracellular domain of ALK7. In additional embodiments, the ALK7-binding protein is an anti-ALK7 antibody such as a full-length anti-ALK7 antibody or an ALK7-binding antibody fragment and variants and derivatives thereof.

[0285] ALK7 binding proteins can be readily prepared using known techniques. Monoclonal anti-ALK7 antibodies can be prepared using techniques known in the art, including hybridoma methods such as those described by Kohler and Milstein, Nature 256:495-497 (1975). Using the hybridoma approach, mice, hamsters, or other suitable host animals are immunized as described above to elicit lymphocytes to produce antibodies that specifically bind to the immunized antigen. Lymphocytes can also be immunized in vitro. Following immunization, lymphocytes are isolated and fused with an appropriate myeloma cell line to form hybridoma cells, which can then be selec...

Embodiment 1

[0404] Example 1. Selection, Characterization and Generation of ALK7 Binding Antibodies

[0405] Human IgG antibodies that bind ALK7 with high affinity were selected using a multi-round selection procedure, which is described in detail below.

[0406] Materials and methods

[0407] Proteins containing human ALK7-Fc were biotinylated using the EZ-Link Sulfur-NHS-Biotinylation kit from Pierce. Goat anti-human F(ab') 2 Kappa-FITC (LC-FITC), Extravidin-PE (EA-PE) and streptavidin-633 (SA-633) were obtained from Southern Biotech, Sigma and Molecular Probes, respectively. Streptavidin microbeads and MACS LC separation column were purchased from Miltenyi Biotec.

[0408] Experiments were performed at 25°C and 37°C using Biacore T100 / T200 biosensors (Biacore / GE Healthcare). ALK7 antibody was captured on a custom FAB chip. A series of concentrations of ALK7-Fc containing protein were injected over the flow cell at a flow rate of 50 μl / ml. To obtain kinetic rate constants, the cor...

Embodiment 2

[0420] Example 2. Characterization of ALK7-binding antibodies

[0421] Exemplary ALK7-binding proteins generated according to the preceding examples were further characterized by sequence, SPR, and cell-based lipolysis inhibition assay analysis.

[0422] The sequences of exemplary ALK7-binding antibodies generated according to the methods described in Example 1 are presented in Table 1A (exemplary CDR sequences are underlined).

[0423] Table 1A : Exemplary ALK7 binding protein

[0424] G04

[0425] VH FR1

QVQLVQSGAEVKKPGSSVKVSCKASGGTFS (SEQ ID NO: 6)

VH CDRI

SYAIS (SEQ ID NO: 1)

VH FR2

WVRQAPGQGLEWMG (SEQ ID NO: 7)

VH CDR2

GIIPIFGTASYAQKFQG (SEQ ID NO: 2)

VH FR3

RVTITADESTSTAYMELSSLRSEDTAVYYCAR (SEQ ID NO: 8)

[0426]

[0427] C02

[0428]

[0429]

[0430] D04

[0431]

[0432] H03

[0433]

[0434]

[0435] Table 1B : Additional Exemplary ALK7 Binding Proteins

[0436] J01

[0437] ...

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Abstract

This disclosure provides ALK7-binding proteins such as anti-ALK7 antibodies, and compositions and methods for making the ALK7-binding proteins. In certain embodiments the ALK7-binding proteins inhibit, or antagonize ALK7 activity. In addition, the disclosure provides compositions and methods for diagnosing and treating overweight, obesity, diabetes, overweight, obesity, type 2 diabetes, and theirassociated conditions; metabolic disorders, and other diseases or conditions that can be treated, prevented or ameliorated by targeting ALK7.

Description

[0001] Citation of Sequence Listings Submitted Electronically [0002] The contents of the electronically filed Sequence Listing in ASCII text file 3174.004PC01_SeqList.ST25txt (size: 72,898 bytes; and date created: April 21, 2017) filed with this application are hereby incorporated by reference in their entirety. Background technique [0003] Overweight and obesity have reached epidemic proportions in the United States and in many countries around the world, increasing in all age, racial and ethnic groups and in both men and women. Overweight and obesity are also associated with other diseases or conditions that disrupt life activities and lifestyles. Obesity is considered a serious risk factor for other diseases and conditions, such as type 2 diabetes, inflammatory and cardiovascular diseases, lung disease, fatty liver disease, neurological disease, and liver and kidney disease. [0004] Type 2 diabetes is a chronic progressive disease that has also reached epidemic proport...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00
CPCC07K16/40A61P3/04A61P3/06A61P1/16A61P3/10A61P9/10A61P9/00A61P9/04A61P9/12A61P25/00A61P27/02A61P25/28A61P11/00A61P13/12A61P1/18A61P15/00A61P19/06A61P35/00C07K2317/56C07K2317/565C07K2317/76C07K2317/92C07K2317/52A61K2039/505C07K16/2863
Inventor J·克诺夫J·贝尔克N·J·夏基R·库马尔A·格林贝格D·萨科R·卡斯顿圭Y·达贡
Owner ACCELERON PHARMA INC
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