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Anti-dll3 chimeric antigen receptors and uses thereof

A chimeric antigen receptor and single domain antibody technology, applied in the field of binding proteins, can solve problems such as intractable treatment

Pending Publication Date: 2022-02-18
NANJING LEGEND BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is because it is highly aggressive, with about two-thirds of patients having metastatic disease at diagnosis, and is extremely refractory to conventional treatments such as platinum-based chemotherapy

Method used

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  • Anti-dll3 chimeric antigen receptors and uses thereof
  • Anti-dll3 chimeric antigen receptors and uses thereof
  • Anti-dll3 chimeric antigen receptors and uses thereof

Examples

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example

[0212] The examples described herein are not intended to imply that all or only these experiments were performed below. Efforts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

example 1

[0213] Example 1. Animal immunization and antibody library construction

[0214] This example demonstrates that immunized camels show good immune response to human or rhesus monkey DLL3 protein and the obtained immune library shows good quality.

[0215] animal immunity

[0216] The extracellular domain (aa27-466) of the human DLL3 protein (AdipoGen, AG-40B-0151) containing the N-terminal FLAG tag or / and the plasmid expressing DLL3 or the cells expressing DLL3 (CHO-K1 / DLL3 and / or or CHO-K1 / EGF4) were mixed with adjuvant or PBS and injected into camels. Typically, camels are vaccinated 2-4 times at intervals of 1 to 2 weeks. After multiple rounds of immunization, immune responses against the target antigen DLL3 were assessed by serum titration with enzyme-linked immunosorbent assay (ELISA) and flow cytometry assays.

[0217] Phage display library construction

[0218] use Reagents Total RNA was extracted from lymphocytes of immunized camels according to the manufacturer's...

example 2

[0219] Example 2. Production of Anti-DLL3 Antibodies

[0220] Anti-DLL3 antibodies provided herein include single domain antibodies (sdAbs) produced from immunized camelids or from human Fab isolated from synthetic human Fab libraries.

[0221] Phage display

[0222] The phage display library was obtained by immunization (comprising the extracellular domain (aa27-466) of human DLL3 protein (AdipoGen, AG-40B-0151) with an N-terminal FLAG tag or / and a plasmid expressing DLL3 or a cell expressing DLL3 ( CHO-K1 / DLL3 or / and CHO-K1 / EGF4) immunogen) sdAb construction obtained. Another human Fab phage display library was synthesized. Two phage libraries were rescued and stored at 4°C after filter sterilization for further use. The two phage libraries mentioned above were used to isolate bound phage using protein-based panning as well as cell-based panning. Using both libraries, at least one round of panning was performed against protein-based and cell-based panning methods until...

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Abstract

Provided herein are anti-DLL3 chimeric antigen receptors (CARs), DLL3 binding proteins and uses of such CARs or DLL3 binding proteins in the treatment of DLL3 associated disorders, such as small cell lung cancer.

Description

technical field [0001] The present invention relates to a chimeric antigen receptor (CAR) targeting DLL3 and a binding protein specific to DLL3. The present invention also relates to nucleic acid sequences encoding these CARs or these binding proteins, modified immune cells expressing these CARs, and their use for treating DLL3-related disorders. Background technique [0002] The development of cellular immunotherapy has provided a promising approach to treat a variety of tumors. One such treatment involves the genetic engineering of immune cells, particularly T cells, to express chimeric antigen receptors (CARs) on the cell surface. Chimeric antigen receptors are proteins that combine the specificity of monoclonal antibodies (mAbs) with the effector functions of T cells in a common format. Once CAR is expressed in T cells, CAR-modified T cells (CAR-T or CAR-T cells) can acquire properties such as antigen-specific recognition, anti-tumor reactivity, and proliferation, thus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C07K16/30A61P35/00
CPCC07K16/18C07K14/7051C12N15/86C12N5/0636A61K39/0011A61P35/00A61P35/02C07K2317/622C07K2317/569C07K2317/24C07K2317/22C07K2317/565C07K2317/94C07K2319/03C07K2319/02C12N2510/00A61K2039/5158C07K2319/00C07K16/28C07K2317/92A61K2039/505C07K2317/55C07K2317/35A61K2239/31A61K39/4631A61K2239/38C12N5/0646C12N5/0638A61K39/464402A61K39/464434A61K39/4611C07K14/70517C07K14/70521A61K38/00C07K14/70596C07K14/71
Inventor 赵涛彭园园唐安王素娟杨帅张望武术郝瑞栋
Owner NANJING LEGEND BIOTECH CO LTD
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