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AuthorHarms, Mirjadc.contributor.author
AuthorGilg, Andreadc.contributor.author
AuthorStändker, Ludgerdc.contributor.author
AuthorBeer, Ambrosdc.contributor.author
AuthorMayer, Benjamindc.contributor.author
AuthorRasche, Volkerdc.contributor.author
AuthorGruber, Christian W.dc.contributor.author
AuthorMünch, Jandc.contributor.author
Date of accession2021-06-10T11:40:15Zdc.date.accessioned
Available in OPARU since2021-06-10T11:40:15Zdc.date.available
Date of first publication2020-09-29dc.date.issued
AbstractC-X-C chemokine receptor type 4 (CXCR4) is involved in several intractable disease processes, including HIV infection, cancer cell metastasis, leukemia cell progression, rheumatoid arthritis, asthma and pulmonary fibrosis. Thus, CXCR4 represents a promising drug target and several CXCR4 antagonizing agents are in preclinical or clinical development. Important parameters in drug lead evaluation are determination of binding affinities to the receptor and assessment of their stability and activity in plasma or blood of animals and humans. Here, we designed a microtiter plate-based CXCR4 antibody competition assay that enables to measure inhibitory concentrations ( IC50 values) and affinity constants ( Ki values) of CXCR4 targeting drugs. The assay is based on the observation that most if not all CXCR4 antagonists compete with binding of the fluorescence-tagged CXCR4 antibody 12G5 to the receptor. We demonstrate that this antibody-competition assay allows a convenient and cheap determination of binding affinities of various CXCR4 antagonists in living cells within just 3 h. Moreover, the assay can be performed in the presence of high concentrations of physiologically relevant body fluids, and thus is a useful readout to evaluate stability (i.e. half-life) of CXCR4 ligands in serum/plasma, and even whole human and mouse blood ex vivo. Thus, this optimized 12G5 antibodycompetition assay allows a robust and convenient determination and calculation of various important pharmacological parameters of CXCR4 receptor-drug interaction and may not only foster future drug development but also animal welfare by reducing the number of experimental animals.dc.description.abstract
Languageendc.language.iso
PublisherUniversität Ulmdc.publisher
LicenseCC BY 4.0 Internationaldc.rights
Link to license texthttps://creativecommons.org/licenses/by/4.0/dc.rights.uri
KeywordBiological techniquesdc.subject
KeywordPharmacologydc.subject
Dewey Decimal GroupDDC 570 / Life sciencesdc.subject.ddc
Dewey Decimal GroupDDC 610 / Medicine & healthdc.subject.ddc
MeSHReceptors, Chemokinedc.subject.mesh
MeSHReceptors, CXCR4dc.subject.mesh
MeSHDrug discoverydc.subject.mesh
TitleMicrotiter plate‑based antibody‑competition assay to determine binding affinities and plasma/blood stability of CXCR4 ligandsdc.title
Resource typeWissenschaftlicher Artikeldc.type
VersionpublishedVersiondc.description.version
DOIhttp://dx.doi.org/10.18725/OPARU-38002dc.identifier.doi
URNhttp://nbn-resolving.de/urn:nbn:de:bsz:289-oparu-38064-6dc.identifier.urn
GNDPharmakologiedc.subject.gnd
GNDRezeptordc.subject.gnd
FacultyMedizinische Fakultätuulm.affiliationGeneral
InstitutionUKU. Institut für Molekulare Virologieuulm.affiliationSpecific
InstitutionKompetenzzentrum "Ulm Peptide Pharmaceuticals (U-PEP)"uulm.affiliationSpecific
InstitutionUKU. Klinik für Nuklearmedizinuulm.affiliationSpecific
InstitutionInstitut für Epidemiologie und Medizinische Biometrieuulm.affiliationSpecific
InstitutionUKU. Klinik für Innere Medizin IIuulm.affiliationSpecific
Peer reviewjauulm.peerReview
DCMI TypeTextuulm.typeDCMI
CategoryPublikationenuulm.category
In cooperation withMedizinische Universität Wienuulm.cooperation
DOI of original publication10.1038/s41598-020-73012-4dc.relation1.doi
Source - Title of sourceScientific Reportssource.title
Source - Place of publicationNature Researchsource.publisher
Source - Volume10source.volume
Source - Year2020source.year
Source - Article number16036source.articleNumber
Source - eISSN2045-2322source.identifier.eissn
FundingDEAL-Projektuulm.funding
Bibliographyuulmuulm.bibliographie
xmlui.metadata.dc.relation.isSupplementedByhttps://static-content.springer.com/esm/art%3A10.1038%2Fs41598-020-73012-4/MediaObjects/41598_2020_73012_MOESM1_ESM.pdfdc.relation.isSupplementedBy


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