A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding.
We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcyto...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096340&type=printable |
| _version_ | 1826587174557974528 |
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| author | Hadassah Sade Claudia Baumgartner Adrian Hugenmatter Ekkehard Moessner Per-Ola Freskgård Jens Niewoehner |
| author_facet | Hadassah Sade Claudia Baumgartner Adrian Hugenmatter Ekkehard Moessner Per-Ola Freskgård Jens Niewoehner |
| author_sort | Hadassah Sade |
| collection | DOAJ |
| description | We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation. |
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| format | Article |
| id | doaj.art-85dba1466f024dcaa52c873ed3f88334 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2025-03-14T16:21:15Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj.art-85dba1466f024dcaa52c873ed3f883342025-02-22T05:33:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0194e9634010.1371/journal.pone.0096340A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding.Hadassah SadeClaudia BaumgartnerAdrian HugenmatterEkkehard MoessnerPer-Ola FreskgårdJens NiewoehnerWe have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096340&type=printable |
| spellingShingle | Hadassah Sade Claudia Baumgartner Adrian Hugenmatter Ekkehard Moessner Per-Ola Freskgård Jens Niewoehner A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. PLoS ONE |
| title | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. |
| title_full | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. |
| title_fullStr | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. |
| title_full_unstemmed | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. |
| title_short | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding. |
| title_sort | human blood brain barrier transcytosis assay reveals antibody transcytosis influenced by ph dependent receptor binding |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096340&type=printable |
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