In vivo binding and retention of CD4-specific DARPin 57.2 in macaques.
<h4>Background</h4>The recently described Designed Ankyrin Repeat Protein (DARPin) technology can produce highly selective ligands to a variety of biological targets at a low production cost.<h4>Methodology/principal findings</h4>To investigate the in vivo use of DARPins for...
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Public Library of Science (PLoS)
2010-08-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012455&type=printable |
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| author | Pavel Pugach Anders Krarup Agegnehu Gettie Marcelo Kuroda James Blanchard Michael Piatak Jeffrey D Lifson Alexandra Trkola Melissa Robbiani |
| author_facet | Pavel Pugach Anders Krarup Agegnehu Gettie Marcelo Kuroda James Blanchard Michael Piatak Jeffrey D Lifson Alexandra Trkola Melissa Robbiani |
| author_sort | Pavel Pugach |
| collection | DOAJ |
| description | <h4>Background</h4>The recently described Designed Ankyrin Repeat Protein (DARPin) technology can produce highly selective ligands to a variety of biological targets at a low production cost.<h4>Methodology/principal findings</h4>To investigate the in vivo use of DARPins for future application to novel anti-HIV strategies, we identified potent CD4-specific DARPins that recognize rhesus CD4 and followed the fate of intravenously injected CD4-specific DARPin 57.2 in rhesus macaques. The human CD4-specific DARPin 57.2 bound macaque CD4(+) cells and exhibited potent inhibitory activity against SIV infection in vitro. DARPin 57.2 or the control E3_5 DARPin was injected into rhesus macaques and the fate of cell-free and cell-bound CD4-specific DARPin was evaluated. DARPin-bound CD4(+) cells were detected in the peripheral blood as early as 30 minutes after the injection, decreasing within 6 hours and being almost undetectable within 24 hours. The amount of DARPin bound was dependent on the amount of DARPin injected. CD4-specific DARPin was also detected on CD4(+) cells in the lymph nodes within 30 minutes, which persisted with similar kinetics to blood. More extensive analysis using blood revealed that DARPin 57.2 bound to all CD4(+) cell types (T cells, monocytes, dendritic cells) in vivo and in vitro with the amount of binding directly proportional to the amount of CD4 on the cell surface. Cell-free DARPins were also detected in the plasma, but were rapidly cleared from circulation.<h4>Conclusions/significance</h4>We demonstrated that the CD4-specific DARPin can rapidly and selectively bind its target cells in vivo, warranting further studies on possible clinical use of the DARPin technology. |
| format | Article |
| id | doaj-art-e473bd1bb43d44538de70200ce9f1ae1 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2010-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-e473bd1bb43d44538de70200ce9f1ae12025-08-20T03:07:40ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-08-0158e1245510.1371/journal.pone.0012455In vivo binding and retention of CD4-specific DARPin 57.2 in macaques.Pavel PugachAnders KrarupAgegnehu GettieMarcelo KurodaJames BlanchardMichael PiatakJeffrey D LifsonAlexandra TrkolaMelissa Robbiani<h4>Background</h4>The recently described Designed Ankyrin Repeat Protein (DARPin) technology can produce highly selective ligands to a variety of biological targets at a low production cost.<h4>Methodology/principal findings</h4>To investigate the in vivo use of DARPins for future application to novel anti-HIV strategies, we identified potent CD4-specific DARPins that recognize rhesus CD4 and followed the fate of intravenously injected CD4-specific DARPin 57.2 in rhesus macaques. The human CD4-specific DARPin 57.2 bound macaque CD4(+) cells and exhibited potent inhibitory activity against SIV infection in vitro. DARPin 57.2 or the control E3_5 DARPin was injected into rhesus macaques and the fate of cell-free and cell-bound CD4-specific DARPin was evaluated. DARPin-bound CD4(+) cells were detected in the peripheral blood as early as 30 minutes after the injection, decreasing within 6 hours and being almost undetectable within 24 hours. The amount of DARPin bound was dependent on the amount of DARPin injected. CD4-specific DARPin was also detected on CD4(+) cells in the lymph nodes within 30 minutes, which persisted with similar kinetics to blood. More extensive analysis using blood revealed that DARPin 57.2 bound to all CD4(+) cell types (T cells, monocytes, dendritic cells) in vivo and in vitro with the amount of binding directly proportional to the amount of CD4 on the cell surface. Cell-free DARPins were also detected in the plasma, but were rapidly cleared from circulation.<h4>Conclusions/significance</h4>We demonstrated that the CD4-specific DARPin can rapidly and selectively bind its target cells in vivo, warranting further studies on possible clinical use of the DARPin technology.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012455&type=printable |
| spellingShingle | Pavel Pugach Anders Krarup Agegnehu Gettie Marcelo Kuroda James Blanchard Michael Piatak Jeffrey D Lifson Alexandra Trkola Melissa Robbiani In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. PLoS ONE |
| title | In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. |
| title_full | In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. |
| title_fullStr | In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. |
| title_full_unstemmed | In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. |
| title_short | In vivo binding and retention of CD4-specific DARPin 57.2 in macaques. |
| title_sort | in vivo binding and retention of cd4 specific darpin 57 2 in macaques |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0012455&type=printable |
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