The single-domain antibodies (sdAb), lack light chains, yet possess unique characteristics providing researchers a myriad of options not readily available with conventional antibodies. In comparison, the sdAbs boast significantly higher stability and are much smaller in size, only 15kd. As such sdAbs have the ability to recognize epitopes that their conventional antibody counterparts are simply unable to access. Thus providing the single domain antibodies with a wide range of biotechnical uses including: structural biological, diagnostic and therapeutic applications.
The single variable binding domain known as VHH is of particular interest and possess a number of unique and highly desirable features. A VHH antibody has the ability to: rapidly penetrate tissues and cross the blood-brain barrier; the VHH antibody is able endure extreme pH and temperature ranges allowing better retention of the native protein structure and their unique conformation in the biologically active state. The small size of the VHH antibody also allows it to be well suited for biosensor applications. These VHH sdAbs can be generated using phase display and produced economically within yeast or bacteria.
Single Domain Antibody (sdAb): A Magic Bullet?
The unique properties of size, stability and solubility for a single domain antibody enables scientist and engineers to advance in the field of cancer research, drug-development and therapy. With variety of ways to use single domain antibodies and the ability to effectively target cancer cells, it’s no surprise that single domain antibodies are on the front lines in the fight against cancer. Below are applications to cancer and the refer:
- In-Vivo Tumor Imaging
- Tumorigenesis Studies
- Effector Domains
- Antagonist (for VEGFR2)
- Structural Biology Applications
- Applications for Diagnostic Procedures
- Single Domain Antibody Development from ProSci
- Off-the-shelf Single Domain Antibodies
In-Vivo Tumor Imaging
SDAbs produces a variety of high resolution imaging applications, such as nuclear imaging conjugated, multiphoton imaging, and Near-infrared(NIR) imaging
Camelid single-domain antibody-fragment engineering for (pre)clinical in vivo molecular imaging applications: adjusting the bullet to its target.
De Vos, J., Devoogdt, N., Lahoutte, T., & Muyldermans, S. (2013).
Targeting tumors with nanobodies for cancer imaging and therapy.
Oliveira, S., Heukers, R., Sornkom, J., Kok, R. J., & van Bergen En Henegouwen, P. M. (2013).
Structurally Defined αMHC-II Nanobody-Drug Conjugates: A Therapeutic and Imaging System for B-Cell Lymphoma
Fang, T., Duarte, J. N., Ling, J., Li, Z., Guzman, J. S., & Ploegh, H. L. (2016).
Multiphoton imaging of tumor biomarkers with conjugates of single-domain antibodies and quantum dots Hafian, H., Sukhanova, A., Turini, M., Chames, P., Baty, D., Pluot, M., Cohen, J. H., Nabiev, I., & Millot, J. M. (2014).
Real-time analysis of epithelial-mesenchymal transition using fluorescent single-domain antibodies
Maier, J., Traenkle, B., & Rothbauer, U. (2015).
Tumorigenesis Studies
SDAbs are good crystallography chaperones which help stabilize GCPRs in structural studies of tumors and they can also be used to screening difficult targets
GPCR-targeting nanobodies: attractive research tools, diagnostics, and therapeutics
Mujic-Delic, A., de Wit, R. H., Verkaar, F., & Smit, M. J. (2014).
Application of single-domain antibodies in tumor histochemistry.
Maik, K. T., & Mackenzie, C. R. (2012).
Conformational biosensors reveal GPCR signalling from endosomes
Irannejad, R., Tomshine, J. C., Tomshine, J. R., Chevalier, M., Mahoney, J. P., Steyaert, J., Rasmussen, S. G., Sunahara, R. K., El-Samad, H., Huang, B., & von Zastrow, M. (2013).
Antibody fragments for stabilization and crystallization of G protein-coupled receptors and their signaling complexes
Shukla, A. K., Gupta, C., Srivastava, A., & Jaiman, D. (2015).
Neutralizing Nanobodies Targeting Diverse Chemokines Effectively Inhibit Chemokine Function
Ghosh, E., Kumari, P., Jaiman, D., & Shukla, A. K. (2015).
Effector Domains
SDAbs can target parts of drug delivery systems such as GPCRs, ERFPS, liposomes, polymeric micelles or albumin nanoparticles and deliver effector domain enzyme or toxin to or into target cell
Nanobody-based cancer therapy of solid tumors
Kijanka, M., Dorresteijn, B., Oliveira, S., & van Bergen en Henegouwen, P. M. (2015).
Combined MUC1-specific nanobody-tagged PEG-polyethylenimine polyplex targeting and transcriptional targeting of tBid transgene for directed killing of MUC1 over-expressing tumour cells
Sadeqzadeh, E., Rahbarizadeh, F., Ahmadvand, D., Rasaee, M. J., Parhamifar, L., & Moghimi, S. M. (2011).
Salmonella engineered to express CD20-targeting antibodies and a drug-converting enzyme can eradicate human lymphomas
Massa, P. E., Paniccia, A., Monegal, A., de Marco, A., & Rescigno, M. (2013).
Development of oligoclonal nanobodies for targeting the tumor-associated glycoprotein 72 antigen
Sharifzadeh, Z., Rahbarizadeh, F., Shokrgozar, M. A., Ahmadvand, D., Mahboudi, F., Rahimi Jamnani, F., & Aghaee Bakhtiari, S. H. (2013).
Antagonist (for VEGFR2)
SDAbs are functional antibodies that can compete with ligands for binding to receptors or bind to the ligands to prevent receptor phosphorylation/activation.
A nanobody directed to a functional epitope on VEGF, as a novel strategy for cancer treatment
Farajpour, Z., Rahbarizadeh, F., Kazemi, B., & Ahmadvand, D. (2014).
A single-step procedure of recombinant library construction for the selection of efficiently produced llama VH binders directed against cancer markers
Kastelic, D., Frkovic-Grazio, S., Baty, D., Truan, G., Komel, R., & Pompon, D. (2009).
Llama-derived single variable domains (nanobodies) directed against chemokine receptor CXCR7 reduce head and neck cancer cell growth in vivo
Maussang, D., Mujic-Delic, A., Descamps, F. J., Stortelers, C., Vanlandschoot, P., Stigter-van Walsum, M., Vischer, H. F., van Roy, M., Vosjan, M., Gonzalez-Pajuelo, M., van Dongen, G. A., Merchiers, P., van Rompaey, P., & Smit, M. J. (2013).
The anti-tumor activity of a neutralizing nanobody targeting leptin receptor in a mouse model of melanoma McMurphy, T., Xiao, R., Magee, D., Slater, A., Zabeau, L., Tavernier, J., & Cao, L. (2014).
Neutralizing Nanobodies Targeting Diverse Chemokines Effectively Inhibit Chemokine Function
Blanchetot, C., Verzijl, D., Mujić-Delić, A., Bosch, L., Rem, L., Leurs, R., … Smit, M. J. (2013).
Structural Biology Applications
Creating diffraction-quality crystals is a bottleneck in macromolecular x-ray crystallography. Single domain antibodies’ size, stability, and functional capabilities, make them ideal chaperones for crystallizing complex biological systems. Single domain antibodies can target membrane proteins, transient multiprotein assemblies, transient conformational states, intrinsically disordered proteins, protein misfolding and fibril formation, and can selectively stabilize conformational states of membrane proteins making them a preferred tool in crystallography.
Applications for Diagnostic Procedures
Single domain antibodies can be used as biosensors to track conformational properties of targets inside of living cells since they can be expressed as intrabodies in eukarotic cells. They can also constrain protein targets in unique disease-linked conformations that can facilitate the discovery of new therapeutic molecules. When fragments from two separate monoclonal antibodies are combined they are known as bispecific monoclonal antibodies, and are capable of binding two unique antigen targets. These artificial proteins are emerging as useful tools, particularly in cancer immunotherapy procedures.
Single Domain Antibody Development
Throughout all five phases (from immunization to production), single domain integrity is ensured with ProSci’s unwavering commitment to customer satisfaction and multiple quality control checks. If your clinical application calls for a single domain antibody (VHH antibody), purchase ProSci single domain antibodies with confidence! ProSci have earned the trust from both private and public research sectors, working in a variety of research applications – therapeutic, general research, diagnostic, pharmaceutical and many more.
Off-the-shelf Single Domain Antibodies
ProSci product are distributed in the UK and Europe by Caltag Medsystems. They offer a range of highly validated single domain antibodies against immune checkpoint targets such as PD-1 and PD-L1. You can learn more about ProSci’s products and services here.
If you have any questions about these products, please contact us.
Originally posted on ProSci-inc.com