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Antibody-dependent cellular cytotoxicity assay

A technology of cytotoxicity and dependence, applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve the problems of inability to distinguish target cells from effector cell death, etc., achieve high-throughput screening of ADCC activity optimization, and easy operation , the effect of high sensitivity

Inactive Publication Date: 2009-03-25
NOVARTIS AG
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the LDH assay is not a real-time assay, and it cannot distinguish target cell death from effector cell death because both types of cells will release LDH during lysis

Method used

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  • Antibody-dependent cellular cytotoxicity assay
  • Antibody-dependent cellular cytotoxicity assay
  • Antibody-dependent cellular cytotoxicity assay

Examples

Experimental program
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Embodiment

[0102] 1. Foreword by ACEA Biosciences (San Diego, CA) The (Real-Time Cell Electronic Sensor) system uses an electronic readout of impedance to non-invasively quantify cell status in real-time. Cells were seeded in E-Plate microplates (ACEA Biosciences) and integrated with microelectrode sensor arrays. The interaction of cells and microelectrode surfaces results in the generation of cell-electrode impedance responses, which indicate the state of cells in terms of morphology, adhesion quality, and quantity.

[0103] ACEA was used to develop a real-time ADCC assay for adherent cells. Two cell lines were used: SKBR3 (a breast cancer cell line); and MG63 (an osteoblast cell line). SKBR3 is an adherent cell line that overexpresses the HER-2 antigen. is a humanized IgG1 antibody against HER-2 that mediates killing of SKBR3 cells via ADCC in the presence of effector cells. MG63 is an adherent cell line that overexpresses M-SCF. Chir-RX1, a humanized antibody against M-CSF, med...

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Abstract

Methods for detecting antibody dependent cellular cytotoxicity (ADCC) are described herein. The methods are label-free, and can be performed in real time on adherent cells. The methods can include, for example, (a) monitoring the impedance between electrodes on a non-conducting substrate that supports the growth of target cells in an assay medium; and (b) adding effector cells and an antibody that binds to the target cells to the assay medium; wherein any decrease in the impedance between the electrodes on the substrate following addition of the effector cells and the antibody is indicative of ADCC function having been effected in the assay medium.

Description

[0001] Cross Reference Related Applications [0002] This application claims the benefit of and priority of US Provisional Application Serial No. 60 / 756,301 filed January 4, 2006. technical field [0003] The present application relates to methods for detecting antibody-dependent cellular cytotoxicity (ADCC) and (in some embodiments) to methods for detecting ADCC in real time that do not require labeling of cells. Background technique [0004] ADCC is a component of the immune response in which IgG antibodies bind to antigens on the surface of pathogenic or tumorigenic target cells, labeling them and destroying them by NK effector cells. Effector cells having Fcγ receptors (FcγR) recognize and bind to the Fc region of the antibody bound to the target cell. Antibodies thus confer specificity on target cell killing mediated by effector cells, forming a bridge between the two cell types and subsequently releasing lytic granules. [0005] Some therapeutic antibodies exert thei...

Claims

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Application Information

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IPC IPC(8): G01N33/50
CPCG01N33/5014
Inventor J·库尼奇刘诚
Owner NOVARTIS AG
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