Yeast platform
The company focuses on yeast molecular biology technology as its research and development direction, and relies on independent innovative technology platforms such as "high-throughput yeast interaction technology", "yeast secretion expression", and "yeast display" to provide a full chain and scenario application solution for yeast
Peptides are synthetically accessible, amenable to chemical tailoring, and have the potential to bind the typically shallow surfaces seen in therapeutically relevant—and historically intractable—protein–protein interactions (PPIs). Peptides can either act as natural ligands in the form of cofactors, coenzymes, and hormones, or directly interact with macromolecules including proteins, RNA, or DNA.
Gene-centered Y1H (Nuclear System)
Yeast one-hybrid system is a technique developed from yeast two-hybrid to study DNA-protein interactions, and is widely used to study the regulation of gene expression in eukaryotic cells, such as identifying DNA binding sites to discover potential binding protein genes, analyzing DNA binding structural domain information, etc.
TF-Centered Y1H (Nuclear System)
A technology based on yeast single hybridization was established to identify the recognition elements of transcription factors, named TF centered YIH. This technology can accurately, quickly, simply, and efficiently identify transcription factor recognition elements, and has broad application prospects in the study of protein DNA element interactions.
DUAL membrane system designed to identify interactions involving integral membrane proteins or membrane-associated proteins is derived from Dualsystems Biotech AG.
Abiotic-Stress Resistance Gene Screening
Saccharomyces cerevisiae belongs to eukaryote, which is closer to the expression system of plants and animals. This expression system includes the following processes: glycosylation, disulfide bond formation, and Post-translational modification of protein folding.
The yeast three-hybrid system is mainly used to study more complex macromolecular interactions including three components, providing a new method for the study of protein-protein-protein, protein-RNA-protein, and protein-small molecule-protein interactions.
The yeast two-hybrid system is a research method for identifying and detecting protein interactions in living cells, which is performed in the eukaryotic model organism yeast, and is now used in several research fields because of its high sensitivity and wide applicability.
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.
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.
Intrabody Selection and Validation
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
VHH for Immunofluorescence
Our Ubiquitin Selectors with Nanotag®
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
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
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
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
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
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
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
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.
FAQ
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Contact Us
Tel: +86 025-85205672
Email: info@pronetbio.com
Room 1001, Floor 10, Building 3C, Nanjing Xianlin Zhigu, No. 6 Qimin West Road, Xianlin Street, Qixia District, NanJing, Jiangsu, China, 210033
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