GlycoCLICK™-based Antibody Labeling Service
Unlocking Precision with GlycoCLICK™-based Antibody Labeling
Clicking chemistry on an antibody is a chemical modification strategy that enables covalent binding to molecules on the surface of target cells by introducing specific chemical functional groups on the antibody molecule. This approach can be used to alter the affinity, stability, and activity of an antibody, thereby enhancing its effect on the target cell. At CD BioGlyco, our GlycoCLICK™-based Labeling Service works by modifying the glycan chains of the biomolecules, including antibodies, to reveal functional groups that can dock with the labeled object. The labeled object is then attached to the biomolecules via the click reaction. The process of GlycoCLICK™-based antibody labeling enables labeling at the molecular level while minimizing non-specific labeling and maintaining the functional integrity of the antibody.
- Firstly, the clickable linker (AbC) needs to be prepared. We use linkers containing tetrazolium (Tz) or methyltetrazolium (MTz) and linkers containing bicyclononyne (BCN). These linkers are attached to the protein labeling site via a polyethylene glycol (PEG) chain.
- Secondly, we perform a reaction to the antibody with AbC so that the linker covalently binds to the antibody.
- The labeled antibody is then conjugated to the antigen on the surface of the target cell, causing the antibody to undergo a cross-linking reaction on the cell surface.
- Cross-linked antibodies form aggregates on the cell surface and are taken up by the cell through endocytosis. Crosslinked antibodies are more efficiently taken up back into the cell.
Typically, we achieve antibody labeling and cross-linking through this process, which alters the biophysical properties of the antibody, enhances its function on the target cell, and enables more efficient load delivery and synthetic modulation of cellular signals.
Publication Data
Technology: SDS-PAGE, immunostaining, and LC-MS
Journal: Journal of the American Chemical Society
IF: 16.383
Published: 2020
Results: This paper alters the physical properties of antibodies on cell surfaces by promoting intramolecular cross-linking of antibodies. This cross-linking can alter the behavior of the antibody by increasing the effective concentration, including increasing the rate of endocytosis and modulating cell signaling pathways. The generalizability of this approach will allow for a variety of future applications, such as increasing the effect of antibodies in load delivery and modifying intracellular signaling pathways of interest using rationally designed synthetic biology tools. This paper utilizes copper-free click reactions, such as the reverse electron demand Diels-Alder reaction, to selectively and rapidly form covalent bonds between molecules. The researchers designed a series of molecules called AbCs, which include Tz/ MTz and BCNs, to be attached to a protein labeling site (N-hydroxysuccinimide ester) via PEG linkers of different lengths. The researchers found that linkers of appropriate lengths help to achieve proper interactions within AbCs when the antibody binds to cell surface antigens. In addition, the reaction rate should be in the appropriate range such that the desired click reaction is slow enough in dilute extracellular media but fast enough to trigger the cross-linking reaction at increased effective concentrations on the cell surface.
Fig.1 The strategy of antibody cross-linking on the cell surface. (Komatsu, et al., 2020)
Applications
- GlycoCLICK™-based antibody labeling can be used to detect the expression and localization of specific proteins in tissue samples. By combining the fluorescent dye with the antibody, the expression level and distribution of the target protein can be directly observed and quantified under the microscope.
- It can be used to detect and locate the presence and expression of specific proteins at the cellular level. By binding a fluorescent dye to an antibody, the location and amount of the target protein can be observed and quantified within the cell.
- GlycoCLICK™-based antibody labeling can be used for the analysis of the presence of specific proteins in protein samples.
Advantages
- GlycoCLICK™-based antibody labeling facilitates precise quantitative analysis of the labels through fluorescence intensity or other measurement methods, enabling the acquisition of accurate and reliable quantitative data.
- Our labeled antibody products are relatively stable and not easily inactivated or degraded. This makes antibodies much more convenient in terms of storage and transportation.
- Our technology exhibits a high degree of specificity, thereby minimizing interference from non-specific background signals and enhancing detection sensitivity.
CD BioGlyco possesses state-of-the-art technologies and a team of highly skilled researchers, dedicated to delivering exceptional GlycoCLICK™-based antibody labeling services that consistently meet the needs and expectations of our esteemed clients. If you have any questions about our services, please feel free to contact us.
Reference
- Komatsu, T.; et al. Antibody clicking as a strategy to modify antibody functionalities on the surface of targeted cells. Journal of the American Chemical Society. 2020, 142(37): 15644-15648.
For research use only. Not intended for any clinical use.
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