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Diagnostic assays to detect tumor antigens in cancer patients

A diagnostic and tumor technology, applied in the direction of receptor/cell surface antigen/cell surface determinant, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, etc. Can solve problems such as reliable detection of tumor cells

Pending Publication Date: 2021-02-26
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity and / or specificity of such assays may not always be sufficient to reliably detect tumor cells, for example in the context of MHC-presented protein-derived peptides

Method used

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  • Diagnostic assays to detect tumor antigens in cancer patients
  • Diagnostic assays to detect tumor antigens in cancer patients
  • Diagnostic assays to detect tumor antigens in cancer patients

Examples

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Effect test

Embodiment 1

[0282] This paper describes a Jurkat NFAT T cell reporter assay using CD20-expressing SUDHDL4 tumor cells as target cells and sorted single clones of Jurkat NFAT T cells expressing anti-CD20-Fab-CD28ATD-CD28CSD-CD3zSSD ( Figure 4 ). As a positive control, certain wells of a 96-well plate (Cellstar Greiner-bio-one, CAT-No. 655185) were treated with 10 μg / ml CD3 antibody (from ) coating at 4°C overnight or at 37°C for at least 1 hour. CD3 antibody-coated wells were washed twice with PBS, which was completely removed after the final wash step. Jurkat NFAT wild-type cells or Jurkat NFAT CAR cells (further referred to as reporter cells) engineered to express the antigen-binding receptor anti-CD20-Fab-CD28ATD-CD28CSD-CD3zSSD were counted and examined for viability using Cedex HiRes. Adjust the cell number to 1x10 per ml 6 living cells. Therefore, an aliquot of the appropriate cell suspension was pelleted at 210 g for 5 minutes at room temperature (RT) and then resuspended in f...

Embodiment 2

[0286] This paper describes the use of SUDHDL4 tumor cells expressing CD20 as target cells and sorted cell populations of Jurkat NFATT cells expressing anti-CD20-Fab-CD28ATD-CD28CSD-CD3zSSD or anti-CD20-crossFab-CD28ATD-CD28CSD-CD3zSSD The Jurkat NFAT T cell reporter gene assay ( Figure 5 ). As a positive control, wells of a 96-well plate (Cellstar Greiner-bio-one, CAT-No. 655185) were treated with 10 μg / ml CD3 antibody (from ) coating, overnight at 4°C. CD3 antibody-coated wells were washed twice with PBS, which was completely removed after the final wash step. Jurkat NFAT wild-type cells or Jurkat NFAT T cells engineered to express anti-CD20-Fab-CD28ATD-CD28CSD-CD3zSSD or anti-CD20-crossFab-CD28ATD-CD28CSD-CD3zSSD (further referred to as reporter cells) were counted using Cedex HiRes and check its liveness. Adjust the cell number to 1x10 per ml 6 living cells. Therefore, an aliquot of the appropriate cell suspension was pelleted at 210 g for 5 minutes at room tempera...

Embodiment 3

[0290] This article describes a Jurkat NFAT T cell reporter assay using CD20-expressing SUDHDL4 tumor cells as target cells and a sorted population of Jurkat NFAT T cells expressing anti-CD20-scFab-CD28ATD-CD28CSD-CD3zSSD ( Figure 6 ).

[0291] As a positive control, wells of a 96-well plate (Cellstar Greiner-bio-one, CAT-No. 655185) were treated with 10 μg / ml CD3 antibody (from ) coating at 4°C overnight or at 37°C for at least 1 hour. CD3 antibody-coated wells were washed twice with PBS, which was completely removed after the final wash step. Jurkat NFAT wild-type T cells or Jurkat NFAT T cells (further referred to as reporter cells) engineered to express anti-CD20-scFab-CD28ATD-CD28CSD-CD3zSSD were counted and examined for viability using Cedex HiRes. Adjust the cell number to 1x10 per ml 6living cells. Therefore, an aliquot of the appropriate cell suspension was pelleted at 210 g for 5 minutes at room temperature (RT) and then resuspended in fresh RPMI-160 + 10% FCS ...

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Abstract

The present invention generally relates to diagnostic assays using cell cultures, in particular to chimeric antigen receptor (CAR) expressing reporter cell assays to analyze samples, in particular patient samples, to diagnose cancer by quantifying the expression of tumor antigens and / or predicting clinical response to cancer immunotherapies. A further aspect of the present invention is to improvesafety of e.g., cancer immunotherapies.

Description

technical field [0001] The present invention relates generally to diagnostic assays using cell cultures, in particular assays for the analysis of chimeric antigen receptors (CARs) expressing reporter cells in samples, particularly patient samples, to quantify the expression of tumor antigens by and / or predict clinical response to cancer immunotherapy to diagnose cancer. Another aspect of the invention is to improve the safety of eg cancer immunotherapy. Background technique [0002] Cancer is one of the leading causes of death in people of all ages. Cancer is an abnormal stage of cells that results in the uncontrolled proliferation of one or more cell populations. Ultimately, proliferation leads to abnormalities in normal biological function, resulting in a variety of clinical and nonclinical symptoms. Tumor cells often exhibit one or several properties that distinguish tumor cells from normal cells, such as morphology, expression of fetal antigens, lack of contact inhibi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50C07K14/725C07K16/28G01N33/68
CPCG01N33/505G01N33/6845C07K14/7051C07K16/2887C07K16/3007C07K16/3038C07K2317/55C07K2317/622C07K2319/03C07K2319/033A61K39/464453A61K39/4631A61K2239/13A61K39/464424A61K39/4611G01N33/57484C40B30/04
Inventor D·黛安娜C·L·S·德隆L·J·哈尼施C·约斯特C·克雷恩E·莫斯纳V·普尔科徐伟
Owner F HOFFMANN LA ROCHE & CO AG
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