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Recombinant VhH Antibodies

Realize all the advantages of recombinant VhH antibodies produced by the experienced team at antibodies-online and Rockland.

We offer an all-inclusive recombinant VhH antibody production service that generates antibodies suitable for a wide range of applications, including cryptic epitope detection, hapten detection, enzyme active site binding, diagnostic imaging, virus neutralization and more.

Our recombinant VhH antibodies are produced using hyperimmunized camelids and single-cell selection from clonal display, providing the desired sensitivity, specificity, and affinity and enabling production in bacterial or mammalian expression systems.

Advantages of VhH Antibodies

Recombinant VhH antibodies, also called nanobodies, are the antigen binding fragment (Fig1: VhH domain, orange) of naturally occurring heavy chain-only antibodies (HCAbs) that are made by hyperimmunized camelids. They can be engineered into monomeric or multimeric, multi-specific formats that possess high target specificity, nanomolar affinity, and a low tendency to aggregate. They are also less prone to the steric hindrance problems that can challenge conventional antibodies, making recombinant VhH antibodies ideally suited for a range of pre-clinical diagnostic and therapeutic applications.

VhH Antibodies

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Our Service Partner: Rockland

Our Service Partner: Rockland

Benefit from expertise over two generations of scientific advances. Rockland enabling biomolecule detection, characterization and analysis since 1962. 60,000 square feet manufacturing facility produce high-quality antibodies, reagents and custom services for your next project

Benefit from expertise over two generations of scientific advances. Rockland enabling biomolecule detection, characterization and analysis since 1962. 60,000 square feet manufacturing facility produce high-quality antibodies, reagents and custom services for your next project

Our Recombinant VhH Antibody Production Process

Production of recombinant VhH antibodies is typically an 8-month process that includes antigen preparation, camelid immunization, library construction, biopanning and clone isolation, clone validation, and VhH antibody production.

Antigen Preparation

Prior to Month 1

We can use a range of macromolecules as antigens—as long as they can be used under BSL-2 conditions—such as protein, recombinant or purified, synthetic peptide, modified peptide, haptens, whole cell lysate, nucleic acid, lipids, and carbohydrate complexes.

Camelid Immunization

Month 2

We generate VhH antibodies in both llama and alpaca, using the immunization schedule you specify and standard or custom adjuvants. We can provide recommendations prior to library construction based on our decades of experience and monitoring of the antibody-mediated immune response (antisera titration).

VhH Library Construction

Month 3

Upon confirmation of a satisfactory immune response, we generate a yeast display library representing the complete repertoire of the naturally-circulating, antigen-binding VhH antibodies. To ensure that the desired binding properties can be found in the library, we analyze critical parameters such as size and diversity.

Biopanning & Clone Isolation

Month 4-5

To isolate clones that bind to the antigen of interest, we use proprietary affinity selection technologies that we’ve developed over decades of successful projects.

Recombinant VhH Antibody Validation

Month 6-7

Isolated single clones are validated using fit-for-purpose studies that include characterization of specificity (binding to the intended target) and cross-reactivity (binding to unrelated antigens) in the conditions relevant to the applications of interest. We can also perform further custom validation for affinity determination, antibody pair identification, and more.

Recombinant VhH Antibody Production

Month 8

After validation, clones are expressed in a bacterial system at the desired scale, and then purified at levels suitable for binding and functional studies.

We can also express recombinant VhH antibodies in eukaryotic systems for custom antibody formats and/or applications and use custom signal sequences or N- or C-terminal tags when required.

Using Recombinant VhH Antibodies

Our recombinant VhH antibodies can easily be engineered into multispecific and multivalent formats as well as genetic fusions to effector domains (i.e., Fc), enzymes, and more.

VHH Flow Cytometry
Figure 2. Flow cytometry analysis of recombinant EGFR x DIG bi-specific antibodies (BsAb44 and BsAb45) binding to EGFR-positive A431 cells or EGFR-negative HEK293T cells. BsAbs were detected with fluorescence-conjugated streptavidin following incubation with biotinylated DIG or directly using a fluorescence-conjugated anti-His antibody.
VHH Kinetic
Figure 3. Kinetic analysis of the sensograms generated from EGFR x DIGrecombinant VhH bi-specific antibody (BsAb 45) binding to EGFR.
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