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 antibodies for you.

Applications

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

Key benefits

  • Enrichment of selected VHHs in intrabodies thanks to yeast selection
  • 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 find functional intracellular antibodies, Hybrigenics makes a selection from a synthetic VHH library (hs2dAb) by Phage Display coupled with Yeast Two-Hybrid. Find more technical information on Antibody Phage Display Selection and Intrabody Enrichment pages. This double screening favors the enrichment in intrabodies that will be functional in cell because we select antibodies that are expressed in the nucleus of the eukaryotic model organism S.cerevisiae.

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

For more explanations, do not hesitate to contact our teams.

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)

 

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