Nanobody-based Purification
Nanobody-based Purification - Related Videos
08:02
Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
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Cited by 1 •
2025
Endocytic and retrograde transport of proteins from the plasma membrane to the trans-Golgi network is essential to maintaining membrane homeostasis and regulating signaling. Here, we describe a method to image and quantify endocytic transport of transmembrane cargo proteins by live-cell microscopy using derivatized anti-GFP nanobodies in HeLa cells.
05:00
Monitoring the Arrival of GFP-Tagged Surface Proteins to the Trans-Golgi Network Using Functionalized Nanobodies
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2025
This video demonstrates a method to study the uptake and retrograde transport of functionalized nanobodies to understand their intracellular trafficking pathways. By introducing radiolabelled sulfate and utilizing isolation and detection techniques, this method enables the study of endocytic uptake and trans-Golgi network arrival of cargo proteins.
04:29
Bimolecular Complementation Affinity Purification to Isolate Two Interacting Proteins
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2025
In this video, we have demonstrated bimolecular complementation affinity purification, BiCAP, to detect and isolate specific protein dimers using conformation-specific single-domain antibodies.
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...
FAQs
Selectivity comes from the nanobody’s high-affinity recognition of a particular molecular target rather than from nonspecific retention by the solid support. After the target binds, washing removes unrelated sample components while retaining the recognized biomolecule. This separation principle is useful when immunology or infection samples contain many proteins and other potentially interfering materials.
Their small, single-domain architecture can support access to epitopes that may be difficult for larger binding molecules to reach. Combined with their stability, this property can improve the recovery of targets while helping preserve native structure and function. That outcome is especially relevant when purified antigens, proteins, or immune complexes must remain suitable for downstream analysis.
High-affinity binding helps retain the intended target during washing, whereas controlled elution releases it from the immobilized nanobody. The balance between these stages affects both enrichment and the condition of the recovered material. Appropriate control is important when purification supports sensitive assays, structural studies, or functional investigations of host-pathogen molecules.
A typical workflow begins with immobilizing nanobodies on a solid support and introducing the complex sample so the target can bind. The support is then washed to remove nonspecific material, followed by elution under controlled conditions. This sequence separates capture from release and produces an enriched fraction for subsequent immunological or infection-related studies.
The method can enrich several target classes, including viral antigens, bacterial proteins, antibodies, and immune complexes. This breadth allows the same affinity-based strategy to address different host-pathogen questions, from isolating microbial components to concentrating immune-related material. The selected nanobody and its target determine what becomes enriched from the original complex sample.
Enriched material can contribute to sensitive assays, structural studies, diagnostic development, and production of reagents for host-pathogen research. Preserving native structure and function is particularly valuable when the isolated biomolecule must be characterized rather than merely detected. The approach therefore connects sample preparation with both analytical investigation and development of immunological tools.