A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-01-29

AUTHORS

Satiro N De Oliveira, Jiexin Wang, Christine Ryan, Sherie L Morrison, Donald B Kohn, Roger P Hollis

ABSTRACT

BACKGROUND: Chimeric Antigen Receptors (CARs) consist of the antigen-recognition portion of a monoclonal antibody fused to an intracellular signaling domain capable of activating T-cells. CARs displayed on the surface of transduced cells perform non-MHC-restricted antigen recognition and activating intracellular signaling pathways for induction of target cytolysis, cytokine secretion and proliferation. Clinical trials are in progress assessing the use of mature T-lymphocytes transduced with CARs targeting CD19 antigen to treat B-lineage malignancies. CD19 is an attractive target for immunotherapy because of its consistent and specific expression in most of the stages of maturation and malignancies of B-lymphocyte origin, but not on hematopoietic stem cells. Antibodies against the extracellular domain of the CAR molecule (anti-Fab, Fc or idiotype) have been used for detection of CAR expression in research and clinical samples by flow cytometry, but may need development for each construct and present significant background in samples from xenograft models. METHODS: A specific reagent for the detection of anti-CD19 CAR expression was developed, a fusion protein consisting of human CD19 extracellular domains and the Fc region of human IgG1 (CD19sIg). Genes encoding CD19sIg fusion proteins were constructed by fusing either exons 1 to 3 (CD19sIg1-3) or exons 1 to 4 (CD19sIg1-4) of the human CD19 cDNA to a human IgG1Fc fragment. These fusion proteins are intended to work in similar fashion as the MHC Tetramers used for identification of antigen-specific T-cells, and may also have other applications in studies of activation of anti-CD19 CAR bearing cells. The CD19sIg proteins were produced from 293 T cells by stable lentiviral vector transduction and purification from culture medium. RESULTS: ELISA assays using several different monoclonal antibodies to CD19 demonstrated dose-related specific binding by the fusion molecule CD19sIg1-4, but no binding by CD19sIg1-3. Conjugation of the CD19sIg1-4 fusion protein to Alexa Fluor 488 allowed specific and sensitive staining of anti-CD19 CAR-bearing cells for flow cytometry assays, detecting as low as 0.5% of CAR-modified primary cells with minimal background staining. CONCLUSIONS: This fusion molecule is a sensitive reagent for detection of anti-CD19 CAR derived from any monoclonal antibody present in CAR-modified T-cells. More... »

PAGES

23-23

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/1479-5876-11-23

DOI

http://dx.doi.org/10.1186/1479-5876-11-23

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1039797895

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23360526


Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
Incoming Citations Browse incoming citations for this publication using opencitations.net

JSON-LD is the canonical representation for SciGraph data.

TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

[
  {
    "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
    "about": [
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/11", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Medical and Health Sciences", 
        "type": "DefinedTerm"
      }, 
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/1107", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Immunology", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Animals", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Antibodies, Monoclonal", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Antigens, CD19", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Cell Proliferation", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Cell Transplantation", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Dose-Response Relationship, Drug", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Enzyme-Linked Immunosorbent Assay", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Flow Cytometry", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "HEK293 Cells", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Humans", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Immunoglobulin Fragments", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Immunoglobulin G", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Leukocytes, Mononuclear", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Lymphocyte Activation", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Mice", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Mice, Inbred NOD", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Mice, SCID", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Protein Structure, Tertiary", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Receptors, Antigen", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "Recombinant Fusion Proteins", 
        "type": "DefinedTerm"
      }, 
      {
        "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
        "name": "T-Lymphocytes", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "Department of Pediatrics, University of California, Los Angeles (UCLA), 10833 Le Conte Avenue, A2-410 MDCC, MC 175217, Los Angeles, CA 90095-1752, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Pediatrics, University of California, Los Angeles (UCLA), 10833 Le Conte Avenue, A2-410 MDCC, MC 175217, Los Angeles, CA 90095-1752, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "De Oliveira", 
        "givenName": "Satiro N", 
        "id": "sg:person.01236414753.52", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01236414753.52"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Wang", 
        "givenName": "Jiexin", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Ryan", 
        "givenName": "Christine", 
        "id": "sg:person.01157716705.36", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01157716705.36"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Morrison", 
        "givenName": "Sherie L", 
        "id": "sg:person.07531043602.24", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.07531043602.24"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Pediatrics, University of California, Los Angeles (UCLA), 10833 Le Conte Avenue, A2-410 MDCC, MC 175217, Los Angeles, CA 90095-1752, USA", 
            "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Kohn", 
        "givenName": "Donald B", 
        "id": "sg:person.01231045607.82", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01231045607.82"
        ], 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA", 
          "id": "http://www.grid.ac/institutes/grid.19006.3e", 
          "name": [
            "Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Hollis", 
        "givenName": "Roger P", 
        "id": "sg:person.01225555371.59", 
        "sameAs": [
          "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01225555371.59"
        ], 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "sg:pub.10.1038/nbt0997-871", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1006020704", 
          "https://doi.org/10.1038/nbt0997-871"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1186/1479-5876-10-29", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1039407555", 
          "https://doi.org/10.1186/1479-5876-10-29"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2013-01-29", 
    "datePublishedReg": "2013-01-29", 
    "description": "BACKGROUND: Chimeric Antigen Receptors (CARs) consist of the antigen-recognition portion of a monoclonal antibody fused to an intracellular signaling domain capable of activating T-cells. CARs displayed on the surface of transduced cells perform non-MHC-restricted antigen recognition and activating intracellular signaling pathways for induction of target cytolysis, cytokine secretion and proliferation. Clinical trials are in progress assessing the use of mature T-lymphocytes transduced with CARs targeting CD19 antigen to treat B-lineage malignancies. CD19 is an attractive target for immunotherapy because of its consistent and specific expression in most of the stages of maturation and malignancies of B-lymphocyte origin, but not on hematopoietic stem cells. Antibodies against the extracellular domain of the CAR molecule (anti-Fab, Fc or idiotype) have been used for detection of CAR expression in research and clinical samples by flow cytometry, but may need development for each construct and present significant background in samples from xenograft models.\nMETHODS: A specific reagent for the detection of anti-CD19 CAR expression was developed, a fusion protein consisting of human CD19 extracellular domains and the Fc region of human IgG1 (CD19sIg). Genes encoding CD19sIg fusion proteins were constructed by fusing either exons 1 to 3 (CD19sIg1-3) or exons 1 to 4 (CD19sIg1-4) of the human CD19 cDNA to a human IgG1Fc fragment. These fusion proteins are intended to work in similar fashion as the MHC Tetramers used for identification of antigen-specific T-cells, and may also have other applications in studies of activation of anti-CD19 CAR bearing cells. The CD19sIg proteins were produced from 293 T cells by stable lentiviral vector transduction and purification from culture medium.\nRESULTS: ELISA assays using several different monoclonal antibodies to CD19 demonstrated dose-related specific binding by the fusion molecule CD19sIg1-4, but no binding by CD19sIg1-3. Conjugation of the CD19sIg1-4 fusion protein to Alexa Fluor 488 allowed specific and sensitive staining of anti-CD19 CAR-bearing cells for flow cytometry assays, detecting as low as 0.5% of CAR-modified primary cells with minimal background staining.\nCONCLUSIONS: This fusion molecule is a sensitive reagent for detection of anti-CD19 CAR derived from any monoclonal antibody present in CAR-modified T-cells.", 
    "genre": "article", 
    "id": "sg:pub.10.1186/1479-5876-11-23", 
    "inLanguage": "en", 
    "isAccessibleForFree": true, 
    "isFundedItemOf": [
      {
        "id": "sg:grant.2436995", 
        "type": "MonetaryGrant"
      }, 
      {
        "id": "sg:grant.2438693", 
        "type": "MonetaryGrant"
      }
    ], 
    "isPartOf": [
      {
        "id": "sg:journal.1032886", 
        "issn": [
          "1479-5876"
        ], 
        "name": "Journal of Translational Medicine", 
        "publisher": "Springer Nature", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "1", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "11"
      }
    ], 
    "keywords": [
      "monoclonal antibodies", 
      "CD19 chimeric antigen receptor", 
      "CAR expression", 
      "chimeric antigen receptor", 
      "flow cytometry assay", 
      "different monoclonal antibodies", 
      "MHC tetramers", 
      "CD19 CAR", 
      "clinical trials", 
      "cytokine secretion", 
      "lymphocyte origin", 
      "xenograft model", 
      "bearing cells", 
      "Fc fusion protein", 
      "stages of maturation", 
      "hematopoietic stem cells", 
      "extracellular domain", 
      "cytometry assays", 
      "flow cytometry", 
      "lineage malignancies", 
      "fusion protein", 
      "study of activation", 
      "CD19", 
      "ELISA assays", 
      "antigen recognition", 
      "antigen receptor", 
      "antibodies", 
      "exon 1", 
      "clinical samples", 
      "human IgG1", 
      "target cytolysis", 
      "lentiviral vector transduction", 
      "minimal background staining", 
      "malignancy", 
      "fusion molecule", 
      "CAR molecules", 
      "Fc region", 
      "receptor consist", 
      "stem cells", 
      "vector transduction", 
      "specific binding", 
      "primary cells", 
      "attractive target", 
      "staining", 
      "cells", 
      "background staining", 
      "expression", 
      "assays", 
      "Alexa Fluor 488", 
      "immunotherapy", 
      "culture medium", 
      "lymphocytes", 
      "protein", 
      "specific reagents", 
      "IgG1", 
      "cytolysis", 
      "specific expression", 
      "similar fashion", 
      "MHC", 
      "secretion", 
      "trials", 
      "cytometry", 
      "receptors", 
      "intracellular", 
      "proliferation", 
      "induction", 
      "activation", 
      "maturation", 
      "sensitive staining", 
      "pathway", 
      "detection", 
      "samples", 
      "target", 
      "study", 
      "genes", 
      "background", 
      "transduction", 
      "binding", 
      "sensitive reagent", 
      "fashion", 
      "use", 
      "cDNA", 
      "stage", 
      "molecules", 
      "identification", 
      "portion", 
      "development", 
      "conjugation", 
      "origin", 
      "fragments", 
      "progress", 
      "reagents", 
      "recognition", 
      "constructs", 
      "significant background", 
      "domain", 
      "region", 
      "research", 
      "medium", 
      "model", 
      "tetramer", 
      "purification", 
      "car", 
      "consist", 
      "surface", 
      "applications", 
      "Chimeric Antigen Receptors (CARs) consist", 
      "Antigen Receptors (CARs) consist", 
      "antigen-recognition portion", 
      "present significant background", 
      "CD19 CAR expression", 
      "human CD19 extracellular domains", 
      "CD19 extracellular domains", 
      "CD19sIg fusion proteins", 
      "human CD19 cDNA", 
      "CD19 cDNA", 
      "human IgG1Fc fragment", 
      "IgG1Fc fragment", 
      "CD19 CAR bearing cells", 
      "CAR bearing cells", 
      "CD19sIg proteins", 
      "stable lentiviral vector transduction", 
      "dose-related specific binding", 
      "fusion molecule CD19sIg1-4", 
      "molecule CD19sIg1-4", 
      "CD19sIg1-4", 
      "CD19sIg1-3", 
      "CD19sIg1-4 fusion protein", 
      "Fluor 488", 
      "CD19/Fc fusion protein"
    ], 
    "name": "A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors", 
    "pagination": "23-23", 
    "productId": [
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1039797895"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1186/1479-5876-11-23"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "23360526"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1186/1479-5876-11-23", 
      "https://app.dimensions.ai/details/publication/pub.1039797895"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2021-12-01T19:29", 
    "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
    "sdPublisher": {
      "name": "Springer Nature - SN SciGraph project", 
      "type": "Organization"
    }, 
    "sdSource": "s3://com-springernature-scigraph/baseset/20211201/entities/gbq_results/article/article_607.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "https://doi.org/10.1186/1479-5876-11-23"
  }
]
 

Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1186/1479-5876-11-23'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1186/1479-5876-11-23'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1186/1479-5876-11-23'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1186/1479-5876-11-23'


 

This table displays all metadata directly associated to this object as RDF triples.

323 TRIPLES      22 PREDICATES      179 URIs      169 LITERALS      28 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1186/1479-5876-11-23 schema:about N1ba8613298244c2a9b96753cb28a645e
2 N2ca722fd0507458abf29b10d8fcf9e91
3 N5c0ccb4386d14cc38a09bf37694cc5ce
4 N73d88a02af424e0c8c2113811210eaf2
5 N80fc1dee00ce4860bad4db4a41edb94b
6 N8f17ca8e4b3843e587bd5ec19eb7c19b
7 N8f4cd895d69e435d841e5aeab8c3e7bb
8 N903ad1186a5c4897aeed76b674b137d6
9 N90d38902cfee4c389d3404235c09b979
10 N9155efca83be4193b7574aed56765f2c
11 N97e6ced00a214e93942d593c4e28de82
12 Na824ec450e1942faabcdabde18c37dc2
13 Naf35bac9437a4d1396d5abc8a23be9f2
14 Nb14ad62699ea4f58b2806f4fdd8f5585
15 Nb68743508cea405f9d62dcd5882d41a9
16 Nc2dc6c94c1524858946c1781ce34a037
17 Nd2045b494a5d424f93270f6b86b1e1fc
18 Ne643971a959f4d329cfa318c00937bfa
19 Nec9f2ac5b2b74161864da160a22957ab
20 Ned80ce6664594bbe820382fcac961b57
21 Nf5d18123e4e64b869ff1d68c1efa4e3c
22 anzsrc-for:11
23 anzsrc-for:1107
24 schema:author N3abd7ba497ae42e1afcf37559e437251
25 schema:citation sg:pub.10.1038/nbt0997-871
26 sg:pub.10.1186/1479-5876-10-29
27 schema:datePublished 2013-01-29
28 schema:datePublishedReg 2013-01-29
29 schema:description BACKGROUND: Chimeric Antigen Receptors (CARs) consist of the antigen-recognition portion of a monoclonal antibody fused to an intracellular signaling domain capable of activating T-cells. CARs displayed on the surface of transduced cells perform non-MHC-restricted antigen recognition and activating intracellular signaling pathways for induction of target cytolysis, cytokine secretion and proliferation. Clinical trials are in progress assessing the use of mature T-lymphocytes transduced with CARs targeting CD19 antigen to treat B-lineage malignancies. CD19 is an attractive target for immunotherapy because of its consistent and specific expression in most of the stages of maturation and malignancies of B-lymphocyte origin, but not on hematopoietic stem cells. Antibodies against the extracellular domain of the CAR molecule (anti-Fab, Fc or idiotype) have been used for detection of CAR expression in research and clinical samples by flow cytometry, but may need development for each construct and present significant background in samples from xenograft models. METHODS: A specific reagent for the detection of anti-CD19 CAR expression was developed, a fusion protein consisting of human CD19 extracellular domains and the Fc region of human IgG1 (CD19sIg). Genes encoding CD19sIg fusion proteins were constructed by fusing either exons 1 to 3 (CD19sIg1-3) or exons 1 to 4 (CD19sIg1-4) of the human CD19 cDNA to a human IgG1Fc fragment. These fusion proteins are intended to work in similar fashion as the MHC Tetramers used for identification of antigen-specific T-cells, and may also have other applications in studies of activation of anti-CD19 CAR bearing cells. The CD19sIg proteins were produced from 293 T cells by stable lentiviral vector transduction and purification from culture medium. RESULTS: ELISA assays using several different monoclonal antibodies to CD19 demonstrated dose-related specific binding by the fusion molecule CD19sIg1-4, but no binding by CD19sIg1-3. Conjugation of the CD19sIg1-4 fusion protein to Alexa Fluor 488 allowed specific and sensitive staining of anti-CD19 CAR-bearing cells for flow cytometry assays, detecting as low as 0.5% of CAR-modified primary cells with minimal background staining. CONCLUSIONS: This fusion molecule is a sensitive reagent for detection of anti-CD19 CAR derived from any monoclonal antibody present in CAR-modified T-cells.
30 schema:genre article
31 schema:inLanguage en
32 schema:isAccessibleForFree true
33 schema:isPartOf N417be8acd2074f2e86edcc8c14e84b49
34 Neeea1df1d0054962972cdd2d5b87e30b
35 sg:journal.1032886
36 schema:keywords Alexa Fluor 488
37 Antigen Receptors (CARs) consist
38 CAR bearing cells
39 CAR expression
40 CAR molecules
41 CD19
42 CD19 CAR
43 CD19 CAR bearing cells
44 CD19 CAR expression
45 CD19 cDNA
46 CD19 chimeric antigen receptor
47 CD19 extracellular domains
48 CD19/Fc fusion protein
49 CD19sIg fusion proteins
50 CD19sIg proteins
51 CD19sIg1-3
52 CD19sIg1-4
53 CD19sIg1-4 fusion protein
54 Chimeric Antigen Receptors (CARs) consist
55 ELISA assays
56 Fc fusion protein
57 Fc region
58 Fluor 488
59 IgG1
60 IgG1Fc fragment
61 MHC
62 MHC tetramers
63 activation
64 antibodies
65 antigen receptor
66 antigen recognition
67 antigen-recognition portion
68 applications
69 assays
70 attractive target
71 background
72 background staining
73 bearing cells
74 binding
75 cDNA
76 car
77 cells
78 chimeric antigen receptor
79 clinical samples
80 clinical trials
81 conjugation
82 consist
83 constructs
84 culture medium
85 cytokine secretion
86 cytolysis
87 cytometry
88 cytometry assays
89 detection
90 development
91 different monoclonal antibodies
92 domain
93 dose-related specific binding
94 exon 1
95 expression
96 extracellular domain
97 fashion
98 flow cytometry
99 flow cytometry assay
100 fragments
101 fusion molecule
102 fusion molecule CD19sIg1-4
103 fusion protein
104 genes
105 hematopoietic stem cells
106 human CD19 cDNA
107 human CD19 extracellular domains
108 human IgG1
109 human IgG1Fc fragment
110 identification
111 immunotherapy
112 induction
113 intracellular
114 lentiviral vector transduction
115 lineage malignancies
116 lymphocyte origin
117 lymphocytes
118 malignancy
119 maturation
120 medium
121 minimal background staining
122 model
123 molecule CD19sIg1-4
124 molecules
125 monoclonal antibodies
126 origin
127 pathway
128 portion
129 present significant background
130 primary cells
131 progress
132 proliferation
133 protein
134 purification
135 reagents
136 receptor consist
137 receptors
138 recognition
139 region
140 research
141 samples
142 secretion
143 sensitive reagent
144 sensitive staining
145 significant background
146 similar fashion
147 specific binding
148 specific expression
149 specific reagents
150 stable lentiviral vector transduction
151 stage
152 stages of maturation
153 staining
154 stem cells
155 study
156 study of activation
157 surface
158 target
159 target cytolysis
160 tetramer
161 transduction
162 trials
163 use
164 vector transduction
165 xenograft model
166 schema:name A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors
167 schema:pagination 23-23
168 schema:productId N019708600e0f4d83887115e293902f5e
169 Ne955dc0d454a41198a2598c717514f69
170 Nf16b727e855743218c27449d3d5926af
171 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039797895
172 https://doi.org/10.1186/1479-5876-11-23
173 schema:sdDatePublished 2021-12-01T19:29
174 schema:sdLicense https://scigraph.springernature.com/explorer/license/
175 schema:sdPublisher Nec74cdd1713c4932ade75435f534f29c
176 schema:url https://doi.org/10.1186/1479-5876-11-23
177 sgo:license sg:explorer/license/
178 sgo:sdDataset articles
179 rdf:type schema:ScholarlyArticle
180 N019708600e0f4d83887115e293902f5e schema:name pubmed_id
181 schema:value 23360526
182 rdf:type schema:PropertyValue
183 N0f7449a379ec4bf0bf5ecbb6eb19385b rdf:first sg:person.01157716705.36
184 rdf:rest N9d4e30fa44d34564aeeadce2beb1c80e
185 N1ba8613298244c2a9b96753cb28a645e schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
186 schema:name Mice, SCID
187 rdf:type schema:DefinedTerm
188 N2ca722fd0507458abf29b10d8fcf9e91 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
189 schema:name Animals
190 rdf:type schema:DefinedTerm
191 N3abd7ba497ae42e1afcf37559e437251 rdf:first sg:person.01236414753.52
192 rdf:rest Na6a54f2833dd49bd80dc36157f067aeb
193 N417be8acd2074f2e86edcc8c14e84b49 schema:issueNumber 1
194 rdf:type schema:PublicationIssue
195 N5c0ccb4386d14cc38a09bf37694cc5ce schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
196 schema:name Flow Cytometry
197 rdf:type schema:DefinedTerm
198 N6cb874987f4742cd857593c35396accb rdf:first sg:person.01225555371.59
199 rdf:rest rdf:nil
200 N73d88a02af424e0c8c2113811210eaf2 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
201 schema:name Immunoglobulin Fragments
202 rdf:type schema:DefinedTerm
203 N80fc1dee00ce4860bad4db4a41edb94b schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
204 schema:name HEK293 Cells
205 rdf:type schema:DefinedTerm
206 N8f17ca8e4b3843e587bd5ec19eb7c19b schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
207 schema:name T-Lymphocytes
208 rdf:type schema:DefinedTerm
209 N8f4cd895d69e435d841e5aeab8c3e7bb schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
210 schema:name Protein Structure, Tertiary
211 rdf:type schema:DefinedTerm
212 N903ad1186a5c4897aeed76b674b137d6 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
213 schema:name Cell Transplantation
214 rdf:type schema:DefinedTerm
215 N90d38902cfee4c389d3404235c09b979 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
216 schema:name Lymphocyte Activation
217 rdf:type schema:DefinedTerm
218 N9155efca83be4193b7574aed56765f2c schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
219 schema:name Mice
220 rdf:type schema:DefinedTerm
221 N97e6ced00a214e93942d593c4e28de82 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
222 schema:name Recombinant Fusion Proteins
223 rdf:type schema:DefinedTerm
224 N9d4e30fa44d34564aeeadce2beb1c80e rdf:first sg:person.07531043602.24
225 rdf:rest Ne20b7a77b14e4fc5b543c35993e1b312
226 Na6a54f2833dd49bd80dc36157f067aeb rdf:first Ncf033841255b45eb9aa1aea1fa88593a
227 rdf:rest N0f7449a379ec4bf0bf5ecbb6eb19385b
228 Na824ec450e1942faabcdabde18c37dc2 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
229 schema:name Antigens, CD19
230 rdf:type schema:DefinedTerm
231 Naf35bac9437a4d1396d5abc8a23be9f2 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
232 schema:name Receptors, Antigen
233 rdf:type schema:DefinedTerm
234 Nb14ad62699ea4f58b2806f4fdd8f5585 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
235 schema:name Immunoglobulin G
236 rdf:type schema:DefinedTerm
237 Nb68743508cea405f9d62dcd5882d41a9 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
238 schema:name Dose-Response Relationship, Drug
239 rdf:type schema:DefinedTerm
240 Nc2dc6c94c1524858946c1781ce34a037 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
241 schema:name Antibodies, Monoclonal
242 rdf:type schema:DefinedTerm
243 Ncf033841255b45eb9aa1aea1fa88593a schema:affiliation grid-institutes:grid.19006.3e
244 schema:familyName Wang
245 schema:givenName Jiexin
246 rdf:type schema:Person
247 Nd2045b494a5d424f93270f6b86b1e1fc schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
248 schema:name Cell Proliferation
249 rdf:type schema:DefinedTerm
250 Ne20b7a77b14e4fc5b543c35993e1b312 rdf:first sg:person.01231045607.82
251 rdf:rest N6cb874987f4742cd857593c35396accb
252 Ne643971a959f4d329cfa318c00937bfa schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
253 schema:name Mice, Inbred NOD
254 rdf:type schema:DefinedTerm
255 Ne955dc0d454a41198a2598c717514f69 schema:name dimensions_id
256 schema:value pub.1039797895
257 rdf:type schema:PropertyValue
258 Nec74cdd1713c4932ade75435f534f29c schema:name Springer Nature - SN SciGraph project
259 rdf:type schema:Organization
260 Nec9f2ac5b2b74161864da160a22957ab schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
261 schema:name Leukocytes, Mononuclear
262 rdf:type schema:DefinedTerm
263 Ned80ce6664594bbe820382fcac961b57 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
264 schema:name Enzyme-Linked Immunosorbent Assay
265 rdf:type schema:DefinedTerm
266 Neeea1df1d0054962972cdd2d5b87e30b schema:volumeNumber 11
267 rdf:type schema:PublicationVolume
268 Nf16b727e855743218c27449d3d5926af schema:name doi
269 schema:value 10.1186/1479-5876-11-23
270 rdf:type schema:PropertyValue
271 Nf5d18123e4e64b869ff1d68c1efa4e3c schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
272 schema:name Humans
273 rdf:type schema:DefinedTerm
274 anzsrc-for:11 schema:inDefinedTermSet anzsrc-for:
275 schema:name Medical and Health Sciences
276 rdf:type schema:DefinedTerm
277 anzsrc-for:1107 schema:inDefinedTermSet anzsrc-for:
278 schema:name Immunology
279 rdf:type schema:DefinedTerm
280 sg:grant.2436995 http://pending.schema.org/fundedItem sg:pub.10.1186/1479-5876-11-23
281 rdf:type schema:MonetaryGrant
282 sg:grant.2438693 http://pending.schema.org/fundedItem sg:pub.10.1186/1479-5876-11-23
283 rdf:type schema:MonetaryGrant
284 sg:journal.1032886 schema:issn 1479-5876
285 schema:name Journal of Translational Medicine
286 schema:publisher Springer Nature
287 rdf:type schema:Periodical
288 sg:person.01157716705.36 schema:affiliation grid-institutes:grid.19006.3e
289 schema:familyName Ryan
290 schema:givenName Christine
291 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01157716705.36
292 rdf:type schema:Person
293 sg:person.01225555371.59 schema:affiliation grid-institutes:grid.19006.3e
294 schema:familyName Hollis
295 schema:givenName Roger P
296 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01225555371.59
297 rdf:type schema:Person
298 sg:person.01231045607.82 schema:affiliation grid-institutes:grid.19006.3e
299 schema:familyName Kohn
300 schema:givenName Donald B
301 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01231045607.82
302 rdf:type schema:Person
303 sg:person.01236414753.52 schema:affiliation grid-institutes:grid.19006.3e
304 schema:familyName De Oliveira
305 schema:givenName Satiro N
306 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01236414753.52
307 rdf:type schema:Person
308 sg:person.07531043602.24 schema:affiliation grid-institutes:grid.19006.3e
309 schema:familyName Morrison
310 schema:givenName Sherie L
311 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.07531043602.24
312 rdf:type schema:Person
313 sg:pub.10.1038/nbt0997-871 schema:sameAs https://app.dimensions.ai/details/publication/pub.1006020704
314 https://doi.org/10.1038/nbt0997-871
315 rdf:type schema:CreativeWork
316 sg:pub.10.1186/1479-5876-10-29 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039407555
317 https://doi.org/10.1186/1479-5876-10-29
318 rdf:type schema:CreativeWork
319 grid-institutes:grid.19006.3e schema:alternateName Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA
320 Department of Pediatrics, University of California, Los Angeles (UCLA), 10833 Le Conte Avenue, A2-410 MDCC, MC 175217, Los Angeles, CA 90095-1752, USA
321 schema:name Department of Microbiology, Immunology and Molecular Genetics (MIMG), University of California, Los Angeles (UCLA), 610 Charles E. Young Dr. East, Los Angeles, CA 90095-1489, USA
322 Department of Pediatrics, University of California, Los Angeles (UCLA), 10833 Le Conte Avenue, A2-410 MDCC, MC 175217, Los Angeles, CA 90095-1752, USA
323 rdf:type schema:Organization
 




Preview window. Press ESC to close (or click here)


...