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Basic research produces knowledge and understanding of molecular mechanisms.
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We are pharma's dependable and strategic R&D solutions provider supporting early drug discovery programs.
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Whether you work on basic plant biology or crop sciences we support you in fostering innovation in your scientific research.
Our services
Yeast 2-Hybrid based Protein Interactions screening
Our ULTImate Y2H™ is the most comprehensive yeast two-hybrid system identifying protein protein interactions as well as DNA, RNA and small bioactive molecule interactions.
Identification of Drug-Target Interactions
Benefit from our unique YChemH screening platform to identify the protein targets of your small bioactive molecule and understand its mode of action
Molecular Glues and Targeted Protein Degradation
Identify protein-protein interactions induced by small molecules with our versatile Y2HChem screening platform, a Y2H-based technology.
Hybribody
Benefit from our proprietary synthetic and humanized VHH library for customized Nanobody® selection/validation and affinity maturation projects.
Libraries
Resources
Publications
Explore our latest customer publications for insights, inspiration and how-to techniques.
News
Stay tuned for conference news, our newest customer publications and interesting facts and information around our service platforms.
Glossary and Testimonials
Find scientific insights about the Y2H principle, our cutting-edge technologies for VHH selection, validation and affinity maturation as well as testimonials from our customers.
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Our Nanobody® Solutions

Benefit from our proprietary synthetic and humanized VHH library for customized Nanobody® selection, validation and affinity maturation projects.

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Our Technology

Our  synthetic  single  chain  antibody

VHHs have several unique features including small size (15 kDa), high stability/solubility while retaining high binding affinity to their target antigens. VHHs are composed of 3 hypervariable regions, supporting antigen recognition against diverse targets and separated by 4 framework regions.

Our VHH scaffold is humanized for therapeutic applications reducing the level of immunogenicity. There is no animal use, thus antigens are not processed and VHHs can be developed against non-immunogenic or highly conserved proteins.

Proprietary Nanobody Library

Our  synthetic  humanized VHH  library

Our synthetic antibody library is a naive library with a complexity of 3.109 VHHs enabling to select highly functional conformational nanobodies and intrabodies.

Our Hybribody library possesses more diversity than in a herd of llamas! This is attributed to the rapid advances in synthetic biology and creative scientific efforts of our collaborators Drs. F. Perez and S. Moutel (Curie Institute, Paris) and Dr. A. Olichon (CRCT, Toulouse) (https://elifesciences.org/articles/16228).

 

Hybribody is a completely animal free antibody selection service. Discover here what our Lamas look like.

Our Nanobody® Solutions

Selection of Synthetic VHHs - in vitro

Selection of Synthetic VHHs - in vitro
Nanobodies for Cell Surface Antigens
Intrabody Selection and Validation
Optimization : Extracellular Affinity
Maturation by Yeast Display
Optimization : Intracellular Affinity
Maturation by Y2H
Minibody Cloning and Production
VHH for Immunofluorescence
Our Ubiquitin Selectors with Nanotag®
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Our optimized Phage Display selection technology delivers high quality and fully functional humanized recombinant nanobodies recognizing native antigens. Importantly, the structural integrity of the antigen is maintained throughout the entire VHH selection and validation process. Therefore, our nanobodies are more likely to recognize endogenous antigens for in vitro and in vivo applications, such as immunofluorescence, live cell imaging, protein degradation or blocking protein activity.

Key benefits

  • Conformational antibodies
  • Deliverables of the validated VHH binders include DNA sequences and plasmids
  • Customized selection conditions (buffer, temperature, etc)
  • Animal-free technology
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Our Nanobody® Solutions

Selection of Synthetic VHHs - in vitro

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Our optimized Phage Display selection technology delivers high quality and fully functional humanized recombinant nanobodies recognizing native antigens. Importantly, the structural integrity of the antigen is maintained throughout the entire VHH selection and validation process. Therefore, our nanobodies are more likely to recognize endogenous antigens for in vitro and in vivo applications, such as immunofluorescence, live cell imaging, protein degradation or blocking protein activity.

Key benefits

  • Conformational antibodies
  • Deliverables of the validated VHH binders include DNA sequences and plasmids
  • Customized selection conditions (buffer, temperature, etc)
  • Animal-free technology
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Nanobodies are selected against membrane proteins or cell-surface antigens under native conditions.

The VHH candidates are directly selected on living cells by phage display. In this process, our proprietary library is incubated with the antigen expressed on cells. Subsequently, a washing step with antigen-free cells is performed and 3 to 4 rounds of selection follow.


Selected nanobodies are validated by immunofluorescence (IF) or phage-FACS assays. This selection strategy works very well for microbial antigens, such as expressed by parasites, bacteria or viruses.

Key benefits

  • Variety of conformational nanobodies against a cell-surface target
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Our intrabodies represent functional intracellular nanobodies expressed and properly folded in the cell cytoplasm where they recognize their endogenous target.

To enrich for intrabodies, we combine classical Phage Display Screening with our expertise in Yeast Two-Hybrid screening. These functional nanobodies are expressed inside the cell where they target endogenous antigen.


This combined strategy is useful for VHH selection against soluble antigens that are particularly difficult to express or purify in heterologous expression systems.

Key benefits

  • Enrichment of intracellular targeting nanobodies
  • No batch-to-batch variability
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In vitro affinity maturation of nanobodies and scFv.

We use a combination of mutagenesis and yeast display to greatly improve the binding affinity of your nanobody or scFv. A mutant VHH or scFv library (2-5 x 106) derived from the parental sequence is generated by introducing mutations either across the entire length of the antibody sequence or restricted to the CDR regions and cloned into our proprietary vector for yeast surface expression.

Yeast expressing the mutant library is subjected to several rounds of cell sorting and the concentration of antigen is gradually decreased to select nanobodies with the highest binding affinities. Our yeast display platform controls for the antigen-antibody equilibrium, which is a major advantage compared to phage display.

Key benefits

  • Cost-effective and quick optimization method
  • No drift-effect like in phage display
  • Apparent kD measurements
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Maturation of VHH with our Yeast Two-Hybrid technology to obtain higher binding affinities for specifically intracellular applications (Intrabodization).

This service is divided into two phases: 1) creation of a mutant library generated by mutagenesis of the entire molecule or the CDR regions only 2) perform Y2H selection on the mutant library to identify intrabodies with increased binding affinities. The selection pressure is adjusted during the screening process to enrich for intrabodies with increased binding affinities.

Key benefits

  • Cost-effective and rapid optimization process
  • No-drift effect compared to phage display
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Fuse your VHH with a Fc species of your choice to generate minibodies.

The selected VHH candidates are fused to Fc isotype-specific fragments from either human, mouse, sheep, rabbit, dog and rat to generate a “Minibody". This format is easily produced in CHO or HEK cells and can be used to stain antigens with standard antibody reagents.

Key benefits

  • Versatile & flexible use for a vast array of applications (PLA, ELISA…)
  • No batch-to-batch variability
  • Possible fusion with Fc fragments from human, mouse, sheep, rabbit, dog and rat
  • Enhanced avidity
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To identify VHH candidates for immunofluorence (IF) applications, we express your antigen in cells as a fusion protein to a fluorescent tag which is targeted to a discrete subcellular localization. VHH-derived supernatants from E. coli are tested for co-localization by IF using standard secondary antibody reagents. This method enables the ranking of VHH candidates based on immunofluorescence intensity and co-localization analysis.

Key benefits

  • ELISA & IF-validated VHH nanobody® in a single cDNA clone
  • No variability from batch to batch
  • Versatile tool thanks to the possible fusion with Fc fragments from human, mouse, sheep, rabbit, dog and rat
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Ub-Selectors are the result from our Hybribody technology and are commercially available by Nanotag GmbH. These reagents are a perfect illustration of Hybribody nanobody solutions.

 

In general, ubiquitin is non-immunogenic and ubiquitin-chain specific nanobodies cannot be selected in animals. Therefore, our products represent a new entry point in the ubiquitin world and represent a new class of reagents that now can be developed.

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our publications

Customer Publications

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NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies

Moutel S, Bery N, Bernard V, Keller L, Lemesre E, de Marco A, Ligat L, Rain JC, Favre G, Olichon A, Perez F. Elife 2016 Jul 19;5. pii: e16228

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mNG-tagged fusion proteins and nanobodies to visualize tropomyosins in yeast and mammalian cells

Tomoyuki Hatano, Tzer Chyn Lim, Ingrid Billault-Chaumartin, Anubhav Dhar, Ying Gu, Teresa Massam-Wu, William Scott, Sushmitha Adishesha, Bernardo Chapa-y-Lazo, Luke Springall, Lavanya Sivashanmugam, Masanori Mishima, Sophie G. Martin, Snezhana Oliferenko, Saravanan Palani, Mohan K. Balasubramanian Journal of Cell Science, Toole and Resources,| 23 September 2022

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Single-domain antibodies for functional targeting of the signaling scaffold Shoc2

Jang H, Wilson PG, Sau M, Chawla U, Rodgers DW, Galperin E. Mol Immunol. 2020 Feb;118:110-116

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Structural basis for the dominant or recessive character of GLIALCAM mutations found in leukodystrophies

Elorza-Vidal X, Xicoy-Espaulella E, Pla-Casillanis A, Alonso-Gardón M, Gaitán-Peñas H, Engel-Pizcueta C, Fernández-Recio J, Estévez RHum. Mol Genet. 2020 May 8;29(7):1107-1120

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Development and characterization of single-domain antibodies neutralizing protease nexin-1 as tools to increase thrombin generation

Kawecki C, Aymonnier K, Ferrière S, Venisse L, Arocas V, Boulaftali Y, Christophe OD, Lenting PJ, Bouton MC, Denis CV. J Thromb Haemost. 2020 Sep;18(9):2155-2168

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Role of VAMP7-Dependent Secretion of Reticulon 3 in Neurite Growth.

Wojnacki J, Nola S, Bun P, Cholley B, Filippini F, Pressé MT, Lipecka J, Man Lam S, N'guyen J, Simon A, Ouslimani A, Shui G, Fader CM, Colombo MI, Guerrera IC, Galli T. Cell Rep. 2020 Dec 22;33(12):108536

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Single Domain Antibody Fragments as New Tools for the Detection of Neuronal Tau Protein in Cells and in Mice Studies

Dupré E, Danis C, Arrial A, Hanoulle X, Homa M, Cantrelle FX, Merzougui H, Colin M, Rain JC, Buée L, Landrieu I. ACS Chem Neurosci. 2019 Sep 18;10(9):3997-4006

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DNA G-Quadruplex Recognition In Vitro and in Live Cells by a Structure-Specific Nanobody

Silvia Galli, Larry Melidis, Sean M. Flynn, Dhaval Varshney, Angela Simeone, Jochen Spiegel, Sarah K. Madden, David Tannahill, and Shankar Balasubramanian, J. Am. Chem. Soc. 2022, 144, 50, 23096–23103

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Inhibition of Tau seeding by targeting Tau nucleation core within neurons with a single domain antibody fragment

Clément Danis , Elian Dupré , Orgeta Zejneli , Raphaëlle Caillierez , Alexis Arrial , Séverine Bégard , Justine Mortelecque , Sabiha Eddarkaoui , Anne Loyens , François-Xavier Cantrelle , Xavier Hanoulle , Jean-Christophe Rain , Morvane Colin , Luc Buée, Isabelle Landrieu Mol Ther. 2022 Apr 6;30(4):1484-1499.

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A selection and optimization strategy for single-domain antibodies targeting the PHF6 linear peptide within the Tau intrinsically disordered protein

Justine Mortelecque, Orgeta Zejneli, Séverine Bégard, Margarida C. Simões, Lea ElHajjar, Marine Nguyen, François-Xavier Cantrelle, Xavier Hanoulle, Jean-Christophe Rain, Morvane Colin, Cláudio M. Gomes, Luc Buée, Isabelle Landrieu, Clément Danis, Elian Dupré (2024) J Biol Chem DOI: 10.1016/j.jbc.2024.107163

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Mono- and bi-specific nanobodies targeting the CUB domains of PCPE-1 reduce the proteolytic processing of fibrillar procollagens

Priscillia Lagoutte, Jean-Marie Bourhis, Natacha Mariano, Virginie Gueguen-Chaignon, David Vandroux, Catherine Moali, Sandrine Vadon-Le Goff (2024) J Mol Biol https://doi.org/10.1016/j.jmb.2024.168667

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Development of a V5-tag–directed nanobody and its implementation as an intracellular biosensor of GPCR signaling

Manel Zeghal, Kevin Matte, Angelica Venes, Shivani Patel, Geneviève Laroche,Sabina Sarvan, Monika Joshi, Jean-Christophe Rain, Jean-François Couture, PatrickM. Giguère Journal of Biological Chemistry 20 July 2023, S0021-9258(23)02135-X

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mNG-tagged fusion proteins and nanobodies to visualize tropomyosins in yeast and mammalian cells

Tomoyuki Hatano, Tzer Chyn Lim, Ingrid Billault-Chaumartin, Anubhav Dhar, Ying Gu, Teresa Massam-Wu, William Scott, Sushmitha Adishesha, Bernardo Chapa-y-Lazo, Luke Springall, Lavanya Sivashanmugam, Masanori Mishima, Sophie G. Martin, Snezhana Oliferenko, Saravanan Palani, Mohan K. Balasubramanian Journal of Cell Science, Toole and Resources,| 23 September 2022

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NMDA receptor-targeted enrichment of CaMKIIα improves fear memory

Anthony Chifor, Jeongyoon Choi, Joongkyu Park iScience August 2, 2022 DOI: 10.1016/j.isci.2022.104864

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AZ921 recognizes the endogenous USP7 protein by immunofluorescence in HCT116 cells

Céline Reverdy, Jean-Christophe Rain, (2024) Antibody Reports DOI: https://doi.org/10.24450/journals/abrep.2023.e1433

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Deciphering the crystal structure of a novel nanobody against the NEIL1 DNA glycosylase

Marlo K Thompson, Nidhi Sharma, Andrea Thorn, Aishwarya Prakash (2024) Acta Crystallogr D Struct Biol doi: 10.1107/S205979832400038X.

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Development of a single-domain antibody to target a G-quadruplex located on the hepatitis B virus covalently closed circular DNA genome

Gerardo B. Figueroa, Simmone D'souza, Higor S. Pereira, Gunjan Vasudeva, Sara B. Figueroa, Zachary E. Robinson, Maulik D. Badmalia, Vanessa Meier-Stephenson, Jennifer A. Corcoran, Guido van Marle, Yi Ni, Stephan Urban, Carla S. Coffin, Trushar R. Patel

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Development and Characterization of a NovelNEIL1 Nanobody

Marlo K. Thompson, Mark H. Eggers, DanielleFlores, Israel Valenzuela, Zhengrong Yang, Joel F.Andrews, Tom Johnsten, Aishwarya Prakash

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Library publication

Tissue-specific targeting of DNA nanodevices in a multicellular living organism

Kasturi Chakraborty et al. Elife. 2021 Jul 28;10:e67830

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Library publication

Novel FGFR4-Targeting Single-Domain Antibodies for Multiple Targeted Therapies against Rhabdomyosarcoma

Nagjie Alijaj et al. Cancers (Basel). 2020 Nov 10;12(11):3313.

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Library publication

Uncoupling of dynamin polymerization and GTPase activity revealed by the conformation-specific nanobody dynab

Valentina Galli et al. Elife. 2017 Oct 12;6:e25197

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Library publication

Whole-cell biopanning with a synthetic phage display library of nanobodies enabled the recovery of follicle-stimulating hormone receptor inhibitors

Ronan Crepin et al. Biochem Biophys Res Commun. 2017 Dec 2;493(4):1567-1572.

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Library publication

Nanobodies against surface biomarkers enable the analysis of tumor genetic heterogeneity in uveal melanoma patient-derived xenografts

Ronan Crepin et al. Pigment Cell Melanoma Res. 2017 May;30(3):317-327.

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FAQ

Do you have a question?

For in vitro selection using soluble antigen, we kindly ask for 150-200 μg of purified protein (for a protein of ~100 kDa) in a PBS 10% Glycerol buffer or lyophilized. Of course, we are happy to suggest commercial providers and adapt our selection process to meet your needs.

For proteins larger than 100 kDa, more material is required and we will work with you to develop an appropriate biopanning strategy.

We provide up to 10 validated VHH clones in a bacterial expression vector and their sequences.

Please refer to our original publication in Elife (2016), which describes the first fully synthetic phage display library of a humanized llama single-domain antibody. Various biopanning strategies along with proof-of-concept projects and their applications are described in detail. NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
Moutel S, Bery N, Bernard V, Keller L, Lemesre E, de Marco A, Ligat L, Rain JC, Favre G, Olichon A, Perez F
Elife. 2016 Jul 19;5. pii: e16228

The turnaround for a synthetic nanobody project is ~ 3 months. Depending the nature of the selection strategy and validation steps, the project may take longer to complete.

Yes, of course. We can provide a template for a confidentiality agreement, to be reviewed and adapted to the specifications of your project. You can also propose your own template.Please note that any project discussion and project results are kept confidential.

Yes, you will own the IP rights to the VHH sequences once your project is paid in full.

Improved nanobody using a yeast display systemOne of the aims of the project of our team is to develop low-cost protein therapy for rare bleeding disorders using single domain antibody fragments..

Dr Olivier Christophe
INSERM U1176
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Improved nanobody using a yeast display system

One of the aims of the project of our team is to develop low-cost protein therapy for rare bleeding disorders using single domain antibody fragments (nanobodies). Nanobodies have several advantages, including a low (if any) immunogenicity when applied to humans. Further, their small size (±17 kDa) facilitates their molecular engineering.
We obtained by immunizing a llama with our target of interest, a nanobody that has demonstrated in in vitro and in vivo experiments its ability to improve a rare haemorrhagic phenotype.

Fusions to the C-terminal end of the Aga2p mating adhesion of Saccharomyces cerevisiae has been used in many studies for the selection of affinity reagents by yeast display followed by flow cytometric analysis. Here, thanks to the expertise of Hybrigenics and using an improved yeast display system for the screening of nanobody immune libraries, we have been able to obtain a variant of our nanobody of interest with an affinity improved 50 fold compared to the molecule of origin. This variant has kept all its therapeutic potential and is now one of our lead product in our pipeline of products in pre-clinical stage.

Dr Olivier Christophe
INSERM U1176
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My lab has previously used Hybrigenics services to perform Yeast two-hybrid screens. The data that they provided initiated a new direction in our research. They also used Y2H screening to select nanobodies working as intrabodies against our protein of interest. Hybrigenics team have put a large effort to provide us with several candidate clones. All the clones were provided as plasmids for mammalian expression and went through the initial validation. After a more in-depth validation, we now have reagents that work in several different assays. I am very impressed with the customer service and dedication of the scientific team at Hybrigenics.

Dr. Emilia Galperin
University of Kentucky
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We asked Hybrigenics to select intrabodies against four different proteins involved in vesicular trafficking. They quickly provided us with several candidates per protein, in the form of ready-to-use plasmids for mammalian expression. We already validated some of these intrabodies. Fused to a red fluorescent protein, they will be extremely valuable tools to block our target proteins in live cells.

Dr. Thierry Galli
Jacques Monod Institute
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As part of a collaboration under FP7-HEALTH AgedBrainSYSBio program, Hybrigenics selected and validated anti-Fyn VHH nanobodies working inside cells - intrabodies. Our lab currently explores these very exciting novel tools in primary neurons.

Prof. Michel Simmoneau
Centre for Psychiatry and Neuroscience
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We received many VHH nanobodies from Hybrigenics, selected and validated against Tau and phosho-Tau. The biochemical and functional characterization of those binders is ongoing but we can already say that this service opens new avenues for our research.

Dr. Luc Buée
Jean-Pierre Aubert Research Centre
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My group has been using the services provided by Hybrigenics to perform Y2H screens from Arabidopsis thaliana libraries using viral proteins as baits at several occasions, and has been very satisfied.
Not only Hybrigenics teams have a great expertise in bait design, in performing high-coverage screens, and most importantly in scoring the candidates obtained and pinpointing the technical false-positives - which are invaluable assets, but I also found very enjoyable to work with them.
They are friendly, supportive, and highly competent, and I always appreciated the discussions we had together.
I will for sure never ever perform a yeast two-hybrid screen myself again and I strongly recommend their high-quality services, which are really worth the price.

Moreover, following a very successful yeast two-hybrid screen using one of our viral protein as a bait, our group also got the chance to make use of their newly developed Hybribody service, i.e. synthetic single chains nanobodies. Hybrigenics selected several VHH intrabodies, which they further validated in mammalian cells using fluorescence microscopy. Our lab is now currently validating them in plant cells and the preliminary data we obtained appear very promising. In addition to their use as basic research tools to detect our proteins in living cells, the intrabodies obtained may as well turn useful to inhibit key steps in the viral multiplication process, thus opening new possibilities for designing antiviral strategies.

Dr. Isabelle Jupin
Jacques Monod Institute

Let's discuss your project

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