Crosslinking Antigens
Crosslinking Antigens - Related Videos
03:47
Crosslinking Neuronal Surface Proteins in the Mouse Brain using a Chemical Crosslinking Assay
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2025
This video demonstrates the crosslinking of neuronal surface proteins using Bis-sulfosuccinimidyl suberate or BS3. Isolated and chilled mouse brain is sectioned coronally. The desired region is extracted from the sections, minced, and treated with BS3. BS3 crosslinks surface proteins like GABA receptors, leaving internal proteins unaltered. The reaction is quenched with glycine before processing the samples for further analysis.
11:15
Nanosponge Tunability in Size and Crosslinking Density
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Cited by 1 •
2017
This article describes a process for tuning the size and crosslinking density of covalently crosslinked nanoparticles from linear polyesters containing pendant functionality. By tailoring synthesis parameters (polymer molecular weight, pendant functionality incorporation, and crosslinker equivalents), a desired nanoparticle size and crosslinking density can be achieved for drug delivery applications.
09:06
Preparation of DNA-crosslinked Polyacrylamide Hydrogels
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Cited by 5 •
2014
Our laboratory has developed DNA-crosslinked polyacrylamide hydrogels, a dynamic hydrogel system, to better understand the effects of modulating tissue stiffness on cell function. Here, we provide schematics, descriptions, and protocols to prepare these hydrogels.
06:57
Development of an Antigen-driven Colitis Model to Study Presentation of Antigens by Antigen Presenting Cells to T Cells
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Cited by 9 •
2016
In this antigen-driven colitis model, OT-II CD4+ T cells expressing a red fluorescent protein were adoptively transferred into RAG-/- mice that express a green fluorescent protein in mononuclear phagocytes (MPs). The hosts were challenged with Escherichia coli (E.coli) expressing the ovalbumin protein (OVA) fused to a cyan fluorescent protein (CFP).
05:04
Viral Antigen Microarray Assay to Detect Antibody Isotypes in Serum Against Viral Antigen
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2025
In this video, we demonstrate the process for detecting antibody isotypes in multiple human serum samples using a microarray slide printed with antigenic subtypes of influenza virus strains.
FAQs
Bifunctional crosslinking reagents create covalent bridges because each reagent contains two reactive ends that can react with chemical groups on neighboring protein molecules. When those reactions occur under controlled conditions, separate antigen molecules or an antigen and carrier become physically connected. The resulting linkage can stabilize an association for subsequent immunological analysis, although it may also change molecular accessibility.
Linker choice and reaction conditions determine which molecular features remain available after crosslinking antigens. A reagent can create a stable connection while partially masking or repositioning an epitope, the region recognized by an antibody or immune cell. Conditions therefore need careful control, since altered antigen structure may affect binding properties, antibody recognition, or cellular immune recognition.
Crosslinking antigen molecules to one another and attaching them to carrier proteins serve different experimental purposes. Joining antigens preserves or stabilizes associations among the antigen molecules, whereas coupling to a carrier can improve their use in antibody-production studies. The distinction matters because the selected partner changes the molecular arrangement being examined and may influence how the immune system recognizes the material.
A practical workflow starts by selecting a bifunctional reagent whose reactivity is compatible with the antigen or carrier protein, then choosing controlled reaction conditions. The reagent is allowed to form covalent bridges between neighboring molecules. Afterward, researchers assess whether the crosslinked material retains the intended antigen structure, binding properties, and recognition needed for the study.
Crosslinked antigens are useful in diagnostic assays when a stable immobilized antigen preparation is needed. Covalent joining helps retain the antigen in an assay-associated form, making the material available for the planned immunological measurement. Because immobilization and crosslinking can affect epitope accessibility, assay performance depends on preserving the antibody-recognition features relevant to the assay.
Carrier-protein coupling is particularly relevant when the objective is antibody production. Linking an antigen to a carrier can improve the antigen's suitability for that immunological application, but the modification may also change how epitopes are displayed. Researchers therefore need to consider whether the resulting construct preserves the antibody-recognition properties required for the intended immune response.