The future of
single-domain antibodies



Intrabody Selection & Validation

Phage Display & Yeast Two-Hybrid Selection + Intrabody Analysis Process

Are you looking for a recombinant antibody you can express in mammalian cells? Hybrigenics selects validated intracellular nanobodies for you: intrabodies

Applications

  • Basic research: antigen tracking by live cell imaging e.g. video-microscopy and single molecule localization. Functional knock-out e.g. antigen retention or addressing to the proteosome pathway (degradon)

Key benefits

  • Enrichment of selected VHH nanobodies® in intrabodies thanks to selection inside yeast
  • Antibodies resistant to cytosol reductive environment
  • Antibodies recognizing endogenous proteins 
  • Cost-effective production of VHH intrabodies from delivered cDNA clones
  • No variability from batch to batch
  • Versatile tool to use in a vast array of experiments thanks to the possible fusion with Fc fragments from human, mouse or rabbit (see Minibody Cloning & Production)

How does it work?

To identify functional intracellular nanobodies, Hybrigenics runs a selection from its synthetic VHH library (hs2dAb) using Phage Display coupled with Yeast Two-Hybrid. Find more technical information on Antibody Phage Display Selection and Intrabody Enrichment pages. This combined screening favors the enrichment in intrabodies that will be functional in cell because we select nanobodies that are expressed and properly folded in the eukaryotic model organism S. cerevisiae.

To validate the intracellular expression of nanobodies and the recognition of endogenous antigens, we express both your antigen and pre-selected VHH nanobodies in cells as fusions with fluorescent proteins and detect their colocalization by fluorescence microscopy.

For more information, please contact us.

Material required

For intrabody selection

  • 100 µg of your purified antigen
  • 5 µg of the cDNA of your antigen

For intrabody validation only

  • 5 µg of the cDNA of your antigen

Deliverables

  • Full report containing the selection process description and the fluorescence microscopy data
  • Sequences and cDNA clones of up to 10 intrabodies VHH in the pMCherry vector (mammalian expression)
Nanobody® is a registered trademark of Ablynx
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