Bio-layer interferometry: Difference between revisions

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{{Short description|Optical biosensing technology}}

[[File:Bio-layer interferometry without analyte binding.gif|thumb|Figure 1 - Overview schematic of a Bio-layer interferometry setup|300x300px]][[File:Thin film interference - soap bubble.gif|thumb|265x265px|Figure 2 - The ligand-analyte layer creates an optical path length difference, reflecting incident light in two different patterns]]'''Bio-layer interferometry''' ('''BLI)''') is an optical biosensing technology that analyzes biomolecular interactions in real-time without the need for fluorescent labeling.<ref name="Apiyo_2017">{{Cite book| vauthors = Apiyo D, Schasfoort R, Schuck P, Marquart A, Gedig ET, Karlsson R, Abdiche YN, Eckman Y, Blum SR, Schasfoort RB |title=Handbook of Surface Plasmon Resonance.|date=2017|publisher=Royal Society of Chemistry|isbn=978-1-78801-139-6|oclc=988866146}}</ref> Alongside [[Surfacesurface plasmon resonance|Surface Plasmon Resonance]] (SPR), BLI is one of few widely available [[Label-free quantification|label-free]] biosensing technologies, a detection style that yields more information in less time than traditional processes.<ref>{{cite journal | vauthors = Syahir A, Usui K, Tomizaki KY, Kajikawa K, Mihara H | title = Label and Label-Free Detection Techniques for Protein Microarrays | journal = Microarrays | volume = 4 | issue = 2 | pages = 228–244 | date = April 2015 | pmid = 27600222 | pmc = 4996399 | doi = 10.3390/microarrays4020228 | doi-access = free }}</ref> The technology relies on the phase shift-wavelength correlation created between interference patterns off of two unique surfaces on the tip of a biosensor.<ref name=":1">{{cite journal | vauthors = Müller-Esparza H, Osorio-Valeriano M, Steube N, Thanbichler M, Randau L | title = Bio-Layer Interferometry Analysis of the Target Binding Activity of CRISPR-Cas Effector Complexes | journal = Frontiers in Molecular Biosciences | volume = 7 | pages = 98 | date = 2020-05-27 | pmid = 32528975 | pmc = 7266957 | doi = 10.3389/fmolb.2020.00098 | doi-access = free }}</ref> BLI has significant applications in quantifying binding strength, measuring protein interactions, and identifying properties of reaction kinetics, such as rate constants and reaction rates.<ref>{{cite journal | vauthors = Rich RL, Myszka DG | title = Higher-throughput, label-free, real-time molecular interaction analysis | journal = Analytical Biochemistry | volume = 361 | issue = 1 | pages = 1–6 | date = February 2007 | pmid = 17145039 | doi = 10.1016/j.ab.2006.10.040 }}</ref>

== Method ==

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=== Biosensor type and selection ===

Bio-layer interferometry relies on [[Biosensor|biosensors]] with a fiber optic tip upon which the ligand is immobilized.<ref name="Apiyo_2017" /> The tip is additionally coated with a matrix biocompatible with the target molecule to limit any non-specific binding. For BLI calculations to work, it is necessary to assume that both the fiber optic tip and the bound ligand and analyte act as thin, reflective surfaces.<ref>{{Cite journal| vauthors = Gao S, Zheng X, Wu J |date=2017|title=A biolayer interferometry-based competitive biosensor for rapid and sensitive detection of saxitoxin |journal=Sensors and Actuators B: Chemical|volume=246|pages=169–174|doi=10.1016/j.snb.2017.02.078|bibcode=2017SeAcB.246..169G |issn=0925-4005}}</ref> The biosensors are disposable, resulting in low costs and high commercial availability.<ref>{{cite journal | vauthors = Abdiche Y, Malashock D, Pinkerton A, Pons J | title = Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet | journal = Analytical Biochemistry | volume = 377 | issue = 2 | pages = 209–217 | date = June 2008 | pmid = 18405656 | doi = 10.1016/j.ab.2008.03.035 | doi-access = free }}</ref> Biosensor selection is determined by the desired test results: kinetic analysis, quantitative analysis, or both.<ref>{{cite journal | vauthors = Yu Y, Mitchell S, Lynaugh H, Brown M, Nobrega RP, Zhi X, Sun T, Caffry I, Cao Y, Yang R, Burnina I, Xu Y, Estep P | display-authors = 6 | title = Understanding ForteBio's Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant | journal = Journal of Biomolecular Screening | volume = 21 | issue = 1 | pages = 88–95 | date = January 2016 | pmid = 26442912 | doi = 10.1177/1087057115609564 | pmc = 4708621 | doi-access = free }}</ref> Most commercially available biosensor types will be grouped into one of these three categories by the BLI manufacturer.<ref name="Apiyo_2017" />

== Applications ==