http://rdf.ncbi.nlm.nih.gov/pubchem/patent/KR-102162096-B1
Outgoing Links
| Predicate | Object |
|---|---|
| classificationCPCAdditional | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N2800-80 |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N33-5014 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-86 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-63 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6897 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N9-22 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N33-56966 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N33-505 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/G01N33-5044 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-6897 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N15-63 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N33-50 |
| filingDate | 2017-10-03-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| grantDate | 2020-10-06-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationDate | 2020-10-06-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | KR-102162096-B1 |
| titleOfInvention | Products and systems for improved quantification of ADCC activity |
| abstract | The activity of many therapeutic antibodies is mediated in part by antibody-dependent cell-mediated cytotoxicity (ADCC). Traditional methods of quantifying ADCC activity are labor intensive and have high levels of intrinsic parameters. This is in part due to the use of complex endpoints, which are difficult to standardize, ie the use of major human NK-cells from other donors as cytotoxic and effector cells. These limitations are engineered to express a low affinity Fc receptor, FcγRIIIa (CD16), responsive to ligation of the Fc moiety of an antibody bound to a specific antigen expressed on target cells by activation of the NFAT responsive reporter gene. This can be partially overcome by the use of effector cell lines. A new recombinant effector cell line is described, wherein the firefly luciferase (FL) reporter gene is used in ADCC assays with engineered target cells, relative to engineered effector cell lines expressing NFAT regulated reporter-gene. , Regulated by a novel synthetic chimeric promoter comprising binding sites for NF-AT, AP1, NFkB, and STAT5, conferring improved sensitivity, improved dynamic range, improved tolerance to human serum and reduced incubation time. do. The target cells were engineered to overexpress an unchanged high level of a specific antigen recognized by a therapeutic antibody, and allogeneic control cells were developed in which the gene encoding a specific drug target was invalidated by genome editing. Based on the B-cell line Raji engineered to overexpress an unchanged level of CD20 with the allogeneic control Raji B-cell line, in which the gene encoding CD20 was nullified, the novel target cells had minimal interference from human serum and a high degree of It was used to quantify the ADCC activity of Rituxan with accuracy. In order to establish a target cell line for quantification of ADCC activity of anti-TNFα monoclonal antibodies, site-specific mutagenesis was used to mutate the protease cleavage site of TNFα, so that the target cells express membrane-bound, non-cleavable TNFα. Let it. In order to prevent the uncut TNFα expressed on the surface of the HEK293 target cell, which binds to the TNFαRII receptor presented on the surface of neighboring cells, from causing cell death, and therefore to prevent the establishment of a permanent cell line, the TNFαRII receptor is It was invalidated using genome editing. To develop a method for quantifying ADCC activity of therapeutic antibodies in serum samples from patients with rheumatoid arthritis and Crohn's disease in which the membrane-bound TNFα-expressing target cells and recombinant effector cells described herein are treated with TNFα antagonists. Was used. ErbB2 was overexpressed in both HEK293 and SKBR3 cells, in which the gene encoding erbB2 was inactivated by CRISPR-Cas9 genome editing, homologous control erbB2 negative target cells as a control for quantification of ADCC activity of trastuzumab It has been established to provide target cells and target cells. Recombinant target cells were also established for quantification of the ADCC activity of cetuximab, in which the EGF receptor was overexpressed in HEK293 cells. Native HEK293 EGFR negative cells were used as allogeneic control cells. |
| priorityDate | 2016-10-04-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
Total number of triples: 258.