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User:Whomtao/Bio-layer interferometry

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Overview schematic of a Bio-layer interferometry setup

Bio-layer interferometry (BLI) is a optical biosensing technology that analyzes biomolecular interactions in real-time without the need for fluorescent labeling.[1] Alongside Surface Plasmon Resonance(SPR), BLI is one of few widely available label-free biosensing technologies, a detection style that allows for a higher volume of information to be obtained in a quicker amount of time compared to traditional processes.[2] The technology relies on the phase shift-wavelength correlation created between interference patterns off of two unique surfaces on the tip of a biosensor.[3] BLI has significant applications in quantifying binding strength, measuring protein interactions, and identifying properties of reaction kinetics, such as rate constants and reaction rates.[4]


Table of contents goes here:

Explanation

Applications

Differences between BLI and SPR

Article body[edit]

Method[edit]

Basic[edit]
Biosensor Type and Selection[edit]

Bio-layer interferometry utilizes the interference of two waves of differing intensities to determine the level of interaction between two molecules.

BLI monitors interactions between a ligand, immobilized on either the tip of a biosensor or the assay, and an analyte in solution.

The binding between a ligand immobilized on the biosensor tip surface and an analyte in solution produces an increase in optical thickness at the biosensor tip, which results in a wavelength shift, Δλ (Figure 3), which is a direct measure of the change in thickness of the biological layer. Interactions are measured in real time, providing the ability to monitor binding specificity, rates of association and dissociation, or concentration, with high precision and accuracy.

Only molecules binding to or dissociating from the biosensor can shift the interference pattern and generate a response profile. Unbound molecules, changes in the refractive index of the surrounding medium, or changes in flow rate do not affect the interference pattern. This is a unique characteristic of bio-layer interferometry and extends its capability to perform in crude samples used in applications for protein-protein interactions, quantitation, affinity, and kinetics.

don't forget to mention "dip-and-read)

Applications[edit]

Analyzing Biomolecular Interactions[edit]

A key use of Bio-layer interferometry is to analyze and quantify interactions between sets of biomolecules.[1] This is extremely useful in pharmaceutical research, in which biomolecule-membrane interaction determines characteristics of a given drug. Due to its ability to achieve high-resolution data and high throughput, BLI has been used to identify biophysical properties of lipid bilayers, allowing for an alternative method of study than the traditional in vitro methods currently used (microscopy, electrophoresis).[5] In addition, BLI can be used to study effector complex-target interactions. Where the traditional Electrophoretic Mobility Shift Assay (EMSA) method can be used, BLI can act as a suitable substitute if the provided benefits (label-free, real-time measurements) are desired.[6]

Measuring Biomolecular Kinetics

Bio-layer interferometry can be used to analyze kinetics in biomolecular systems. Interference patterns found in BLI experiments can be used to calculate rate constants and other kinetic data in biomolecular interactions.[7] The (relatively) lower sensitivity of the BLI sensor results in less response to changes in sample composition. As a result, BLI can also be used to investigate allosteric effects on enzyme conformational changes.[8]

Overview schematic of Surface Plasmon Resonance

Distinguishing Characteristics[edit]

BLI and SPR are both dominant technologies in the label-free instruments market.[1] Despite sharing some similarities in concept, there are significant differences between the two techniques. Micro-fluidic SPR relies on a closed architecture to transport samples to a stationary sensor chip. BLI instead utilizes an open system, shaking multiple wells on a plate to transport the sensors to the samples without need for micro-fluidics.[5] Being a closed system, SPR's association and dissociation phases are limited by the technology's design. BLI's open plate design results in association and dissociation length limits determined by sample evaporation instead.[9] SPR is easily reproducible due to its continuous flow microfluidics. BLI's multi well plate design allows for extremely high throughput in one batch. Assay configuration in BLI can, in stable conditions, allow for recovery of samples. Assay configuration in SPR allows for higher sensitivity. As a result, BLI results are often compared to SPR results for validation.[10]

bio-layer interferometry

References[edit]

  1. ^ a b c David., Apiyo, (2017). Handbook of Surface Plasmon Resonance. Royal Society of Chemistry. ISBN 978-1-78801-139-6. OCLC 988866146.{{cite book}}: CS1 maint: extra punctuation (link) CS1 maint: multiple names: authors list (link)
  2. ^ Syahir, Amir; Usui, Kenji; Tomizaki, Kin-ya; Kajikawa, Kotaro; Mihara, Hisakazu (2015-04-24). "Label and Label-Free Detection Techniques for Protein Microarrays". Microarrays. 4 (2): 228–244. doi:10.3390/microarrays4020228. ISSN 2076-3905.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  3. ^ Müller-Esparza, Hanna; Osorio-Valeriano, Manuel; Steube, Niklas; Thanbichler, Martin; Randau, Lennart (2020-05-27). "Bio-Layer Interferometry Analysis of the Target Binding Activity of CRISPR-Cas Effector Complexes". Frontiers in Molecular Biosciences. 7. doi:10.3389/fmolb.2020.00098. ISSN 2296-889X.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  4. ^ Rich, Rebecca L; Myszka, David G (1 February 2007). "Higher-throughput, label-free, real-time molecular interaction analysis". Analytical Biochemistry. 361 (1): 1–6. doi:10.1016/j.ab.2006.10.040. PMID 17145039.
  5. ^ a b Wallner, Jakob; Lhota, Gabriele; Jeschek, Dominik; Mader, Alexander; Vorauer-Uhl, Karola (2013). "Application of Bio-Layer Interferometry for the analysis of protein/liposome interactions". Journal of Pharmaceutical and Biomedical Analysis. 72: 150–154. doi:10.1016/j.jpba.2012.10.008. ISSN 0731-7085.
  6. ^ Müller-Esparza, Hanna; Osorio-Valeriano, Manuel; Steube, Niklas; Thanbichler, Martin; Randau, Lennart (2020-05-27). "Bio-Layer Interferometry Analysis of the Target Binding Activity of CRISPR-Cas Effector Complexes". Frontiers in Molecular Biosciences. 7. doi:10.3389/fmolb.2020.00098. ISSN 2296-889X.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  7. ^ Wilson, Jo Leanna; Scott, Israel M.; McMurry, Jonathan L. (2010). "Optical biosensing: Kinetics of protein A-IGG binding using biolayer interferometry". Biochemistry and Molecular Biology Education. 38 (6): 400–407. doi:10.1002/bmb.20442. ISSN 1470-8175.
  8. ^ Shah, Naman B.; Duncan, Thomas M. (2014-02-18). "Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects". Journal of Visualized Experiments (84). doi:10.3791/51383. ISSN 1940-087X.
  9. ^ Abdiche, Yasmina; Malashock, Dan; Pinkerton, Alanna; Pons, Jaume (2008). "Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet". Analytical Biochemistry. 377 (2): 209–217. doi:10.1016/j.ab.2008.03.035. ISSN 0003-2697.
  10. ^ Yang, Danlin; Singh, Ajit; Wu, Helen; Kroe-Barrett, Rachel (2016). "Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics". Analytical Biochemistry. 508: 78–96. doi:10.1016/j.ab.2016.06.024. ISSN 0003-2697.
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