Antigen Purification
Antigen Purification - Related Videos
12:48
Purification of the Membrane Compartment for Endoplasmic Reticulum-associated Degradation of Exogenous Antigens in Cross-presentation
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Cited by 2 •
2017
The method described here is a new vesicle isolation protocol, which allows for the purification of the cellular compartments where exogenous antigens are processed by endoplasmic reticulum-associated degradation in cross-presentation.
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).
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
06:30
Gel Purification
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2023
Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, washing and eluting from a silica column. DNA is believed to bind to silica in the presence of high salt via a salt bridge. Following binding, DNA is washed of impurities and eluted under low salt conditions disrupting this interaction. This video goes through a step-by-step, generalized procedure for cutting out a band from the gel, gel...
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
Preservation depends on maintaining the antigen’s structural integrity and biological activity while contaminants are removed. The purification workflow therefore combines separation steps rather than relying only on concentration. If processing alters the molecular features recognized by the immune system, the resulting antigen may perform poorly in antibody production, immunoassays, vaccine studies, or host-pathogen interaction analyses.
These operations contribute different separation functions within the workflow. Clarification and filtration help prepare the biological mixture, precipitation supports antigen concentration or separation, and chromatography provides a more selective purification stage. Used together, they progressively reduce contaminants and produce material suitable for downstream immunological analysis.
Affinity methods use specific binding interactions to capture the target antigen from a mixture. This selectivity can distinguish the antigen from other biological components before unwanted material is removed and the target is eluted. Their value is greatest when downstream experiments require reliable antigen identity and sufficient purity for immune-based measurements.
Purity determines how much non-target material accompanies the antigen, while structural integrity affects whether immune-recognized features remain intact. Biological activity indicates whether the preparation retains relevant functional properties. These characteristics directly influence the reliability and reproducibility of antibody production, immunoassays, pathogen detection, vaccine studies, and host-pathogen interaction research.
A typical workflow begins with clarification and filtration of the complex biological mixture, followed by precipitation and one or more chromatography steps. An affinity stage may selectively capture the antigen, after which contaminants are removed and the target is eluted. The sequence is designed to concentrate the antigen while retaining features needed for immunological applications.
Affinity purification is especially relevant when a specific binding interaction can selectively capture the antigen from a complex mixture. It can serve as a focused chromatography stage after preliminary clarification, filtration, or precipitation. Researchers may choose this approach when the final preparation must support sensitive antibody, detection, or immunoassay work with reduced contaminant interference.
Purified antigens support several distinct research goals, including generating antibodies, developing immunoassays, detecting pathogens, and conducting vaccine studies. They also enable analysis of host-pathogen interactions. Across these applications, consistent purity and preservation of immune-recognized features help investigators interpret antigen-specific responses and compare experimental results more reliably.