Content deleted Content added
added Category:Interferometry using HotCat |
mNo edit summary |
||
| (6 intermediate revisions by 6 users not shown) | |||
Line 1:
{{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
== Method ==
Line 11 ⟶ 12:
=== 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 ==
=== Analyzing biomolecular interactions ===
A key use of Bio-layer interferometry is to analyze and quantify interactions between sets of biomolecules.<ref name="Apiyo_2017" /> 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]]).<ref name=":
=== Measuring biomolecular kinetics ===
Bio-layer interferometry can be used to analyze kinetics in biomolecular systems. The benefits that BLI brings provide additional insight into kinetics on top of commonly used endpoint methods like [[ELISA|enzyme-linked immunosorbent assay]] (ELISA).<ref name="Apiyo_2017" /> Interference patterns found in BLI experiments can be used to calculate rate constants and other kinetic data in biomolecular interactions.<ref>{{cite journal | vauthors = Wilson JL, Scott IM, McMurry JL | title = Optical biosensing: Kinetics of protein A-IGG binding using biolayer interferometry | journal = Biochemistry and Molecular Biology Education | volume = 38 | issue = 6 | pages = 400–407 | date = November 2010 | pmid = 21567869 | doi = 10.1002/bmb.20442 | s2cid = 29689214 }}</ref> 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 regulation|allosteric effects]] on enzyme conformational changes.<ref>{{cite journal | vauthors = Shah NB, Duncan TM | title = Bio-layer interferometry for measuring kinetics of protein-protein interactions and allosteric ligand effects | journal = Journal of Visualized Experiments | issue = 84 | pages = e51383 | date = February 2014 | pmid = 24638157 | pmc = 4089413 | doi = 10.3791/51383 }}</ref>
== Distinguishing characteristics ==
BLI and SPR are both dominant technologies in the label-free instruments market.<ref name="Apiyo_2017" /> 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 (Figure 4). BLI instead utilizes an open system, shaking multiple wells on a plate to transport the sensors to the samples without need for [[Microfluidics|micro-fluidics]].<ref name=":
==See also==
| |||