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@ARTICLE{Bier:877918,
      author       = {Bier, Dirk and Schulze, Annette and Holschbach, Marcus and
                      Neumaier, Bernd and Baumann, A.},
      title        = {{D}evelopment and {E}valuation of a {V}ersatile
                      {R}eceptor-{L}igand {B}inding {A}ssay {U}sing {C}ell
                      {M}embrane {P}reparations {E}mbedded in an {A}garose {G}el
                      {M}atrix and {E}valuation with the {H}uman {A}denosine {A}1
                      {R}eceptor},
      journal      = {Assay and drug development technologies},
      volume       = {18},
      number       = {7},
      issn         = {1540-658X},
      address      = {Larchmont, NY},
      publisher    = {Liebert},
      reportid     = {FZJ-2020-02510},
      pages        = {},
      year         = {2020},
      abstract     = {Guanosine-5′-triphosphate (GTP)-binding protein-coupled
                      receptors are the target of up to $40\%$ of prescribed
                      medications worldwide. To evaluate the suitability of novel
                      receptor ligands, frequently elaborate, time-consuming, and
                      expensive receptor-ligand interaction studies have to be
                      carried out. This work describes the development and proof
                      of principle of a rapid, sensitive, and reliable
                      receptor-ligand binding assay. CHO cells were stably
                      transfected with a construct encoding the human A1 adenosine
                      receptor (hA1AR). For ligand binding assays, membranes from
                      these cells were prepared and embedded in low melting point
                      agarose. These “immobilized” samples were incubated with
                      tritiated 8-cyclopentyl-1,3-dipropylxanthine ([3H]DPCPX), a
                      well-established receptor antagonist. The KD and Bmax values
                      as well as kinetic parameters (kon and koff) of
                      receptor-ligand interaction were determined. Unspecific
                      binding of various radiotracers to either the carrier
                      material or the agarose gel matrix was negligible. The
                      dissociation constant (KD) for [3H]DPCPX at the hA1AR was
                      determined by saturation, competition binding, and kinetic
                      experiments. These studies resulted in KD values of
                      ∼3 nM, which is in good accordance with previously
                      published data obtained from conventional receptor-ligand
                      binding assays. The procedure described in this study
                      simplifies classical binding studies to a kit-like assay.
                      The receptors retained their binding properties even when
                      preparations were dried completely. Transport and delivery
                      of the material are conceivable without loss of biological
                      activity. Therefore, other laboratories can perform binding
                      studies without special equipment or the necessity to run a
                      cell culture laboratory and/or to dissect tissue on their
                      own.},
      cin          = {INM-5 / IBI-1},
      ddc          = {540},
      cid          = {I:(DE-Juel1)INM-5-20090406 / I:(DE-Juel1)IBI-1-20200312},
      pnm          = {573 - Neuroimaging (POF3-573)},
      pid          = {G:(DE-HGF)POF3-573},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:32749852},
      UT           = {WOS:000556911700001},
      doi          = {10.1089/adt.2020.991},
      url          = {https://juser.fz-juelich.de/record/877918},
}