Site-directed antibody immobilization techniques for immunosensors
Asta Makaraviciute1, Almira Ramanaviciene
1NanoTechnas-Center of Nanotechnology and Materials Science, Department of Analytical and Environmental Chemistry, Faculty of Chemistry, Vilnius University, Naugarduko Street 24, LT-03225 Vilnius, Lithuania.
Biosensors & Bioelectronics
|August 6, 2013
Summary
Site-directed antibody immobilization enhances immunosensor performance. This review details three key methods: Fc-binding proteins, antibody fragments, and Fc region oxidation for improved sensitivity and stability in biosensor design.
Area of Science:
- Biomedical Engineering
- Biosensor Technology
- Immunochemistry
Background:
- Immunosensor performance relies heavily on antibody immobilization.
- Random immobilization yields satisfactory results, but site-directed methods offer significant improvements.
Purpose of the Study:
- To review conventional site-directed antibody immobilization techniques for immunosensor design.
- To highlight methods that enhance immunosensor sensitivity, regenerability, and stability.
Main Methods:
- Affinity interactions using Fc-binding proteins (Protein A, G).
- Chemical/genetic engineering of antibody fragments for sulfhydryl group attachment.
- Fc region oligosaccharide oxidation for immobilization.
Main Results:
- Site-directed immobilization significantly improves immunosensor sensitivity, especially on planar supports.
- Each method offers distinct advantages and presents unique challenges.
- Proper orientation and activity of immobilized antibodies are crucial.
Conclusions:
- Site-directed immobilization is superior to random methods for advanced immunosensor development.
- Understanding these techniques is vital for optimizing immunosensor design and function.
- Further research into these methods can lead to more robust and sensitive biosensing platforms.
Related Concept Videos
01:20Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
01:20Immunoprecipitation
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
01:33Enzyme-Linked Immunosorbent Assay
In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.