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SpyMask enables combinatorial assembly of bispecific binders.

Driscoll CL, Keeble AH, Howarth MR
Nat Commun. 2024 Mar 16;15(1):2403. doi: 10.1038/s41467-024-46599-9. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
216280pET28a-SnoopTag-AffiHER2-SpyTagExpresses an affibody to HER2 fused to SnoopTag and SpyTag, for bacterial expression
216281pET28a-SnoopTag-SpyTag-(AffiHER2)x3Three affibodies to HER2 linked in a chain for bacterial expression. SnoopTag and SpyTag allow isopeptide bond formation to their cognate Catcher
216282pET28a-SUMO-SpyTag003Expresses SUMO with a C-terminal SpyTag003 in bacterial cells
216283pDEST14-SpyCatcher003-TEVs-SpyTag003DAExpresses SpyCatcher003-TEV site-SpyTag003DA in bacterial cytoplasm, to allow TEV protease to remove SpyTag003DA and thereby uncage SpyCatcher003 reactivity.
216284pDEST14-DoubleCatcherExpresses SpyCatcher003-SpyCatcher003-TEV site-SpyTag003DA in bacterial cytoplasm, to allow stepwise coupling of two unique SpyTag-linked binders
216285pDEST14-DoubleCatcher-H-LockExpresses DoubleCatcher H-lock in bacterial cells. DoubleCatcher H-lock is a variant of DoubleCatcher in which each SpyCatcher003 moiety is separated by an alpha helical linker.
216286pDEST14-DoubleCatcher-⍺-LockExpresses DoubleCatcher-⍺-Lock in bacterial cells. DoubleCatcher-⍺-Lock is a variant of DoubleCatcher in which the orientation of each SpyCatcher003 moiety is locked via an internal disulfide bond.
216287pDEST14-DoubleCatcher-β-LockExpresses DoubleCatcher-β-Lock in bacterial cells. DoubleCatcher-β-Lock is a variant of DoubleCatcher in which the orientation of each SpyCatcher003 moiety is locked via an internal disulfide bond.
216288pDEST14-DoubleCatcher-γ-LockExpresses DoubleCatcher-γ-Lock in bacterial cells. DoubleCatcher-γ-Lock is a variant of DoubleCatcher in which the orientation of each SpyCatcher003 moiety is locked via an internal disulfide bond.
216289pDEST14-DoubleCatcher-δ-LockExpresses DoubleCatcher-δ-Lock in bacterial cells. DoubleCatcher-δ-Lock is a variant of DoubleCatcher in which the orientation of each SpyCatcher003 moiety is locked via an internal disulfide bond.
216290pDEST14-DoubleCatcher-ε-LockExpresses DoubleCatcher-ε-Lock in bacterial cells. DoubleCatcher-ε-Lock is a variant of DoubleCatcher in which the orientation of each SpyCatcher003 moiety is locked via an internal disulfide bond.
216291pET21b-MBPx-MBPx-SpyTag003Expresses tandem MBPx linked to-SpyTag003 in bacterial cells. MBPx is a variant of MBP (A312V I317V Δ172-173 Δ175-176) with enhanced affinity for maltose.
216292pcDNA3.1-Tras heavy chain-GSG-SpyTag003Expresses anti-HER2 Tras Heavy chain-GSG-SpyTag003 in mammalian cells. To be paired with Tras light chain to form the anti-HER2 Tras Fab
216293pcDNA3.1-Tras heavy chain-SpyTag003Expresses anti-HER2 Tras Heavy chain-SpyTag003 in mammalian cells. To be paired with Tras light chain to form the anti-HER2 Tras NoLink Fab
216294pcDNA3.1-Tras light chainExpresses anti-HER2 Tras Light chain in mammalian cells. To be paired with Tras heavy chain to form the anti-HER2 Tras Fab
216295pET28-SpyCatcher002-MBPExpresses SpyCatcher002 linked to maltose-binding protein in bacterial cells
216296pcDNA3.1-39S Heavy chain-SpyTag003Expresses anti-HER2 39S Heavy chain-SpyTag003 in mammalian cells. To be paired with 39S light chain to form the anti-HER2 39S Fab
216297pcDNA3.1-39S Light chainExpresses anti-HER2 39S Light chain in mammalian cells. To be paired with 39S heavy chain to form the anti-HER2 39S Fab
216303pcDNA3.1-MF3958 Heavy chain-SpyTag003Expresses anti-HER2 MF3958 Heavy chain-SpyTag003 in mammalian cells. To be paired with MF3958 light chain to form the anti-HER2 MF3958 Fab
216304pcDNA3.1-MF3958 Light chainExpresses anti-HER2 MF3958 Light chain in mammalian cells. To be paired with MF3958 heavy chain to form the anti-HER2 MF3958 Fab
216307pcDNA3.1-H2-18 Heavy chain-SpyTag003Expresses anti-HER2 H2-18 Heavy chain-SpyTag003 in mammalian cells. To be paired with H2-18 light chain to form the anti-HER2 H2-18 Fab
216308pcDNA3.1-H2-18 Light chainExpresses anti-HER2 H2-18 Light chain in mammalian cells. To be paired with H2-18 heavy chain to form the anti-HER2 H2-18 Fab
216309pcDNA3.1-Pert Heavy chain-SpyTag003Expresses anti-HER2 Pert Heavy chain-SpyTag003 in mammalian cells. To be paired with Pert light chain to form the anti-HER2 Pert Fab (related to Pertuzumab)
216310pcDNA3.1-Pert Light chainExpresses anti-HER2 Pert Light chain in mammalian cells. To be paired with Pert heavy chain to form the anti-HER2 Pert Fab
216312pET28a-nanoHER2-SpyTag003Expresses nanoHER2-SpyTag003 in bacterial cells. nanoHER2 is an anti-HER2 nanobody.

Antibodies from Article