Bio-layer interferometry: Difference between revisions

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=== 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=":213" /> In addition, BLI can be used to study [[Effector (biology)|effector]] complex-target interactions. Where the traditional [[Electrophoretic mobility shift assay|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.<ref name=":1" />[[File:Surface Plasmon Resonance (SPR).jpg|thumb|Figure 4 - Overview schematic of Surface Plasmon Resonance|324x324px]]

=== 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=":213">{{cite journal | vauthors = Wallner J, Lhota G, Jeschek D, Mader A, Vorauer-Uhl K | title = Application of Bio-Layer Interferometry for the analysis of protein/liposome interactions | journal = Journal of Pharmaceutical and Biomedical Analysis | volume = 72 | pages = 150–154 | date = January 2013 | pmid = 23146240 | doi = 10.1016/j.jpba.2012.10.008 }}</ref> 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.<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> 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.<ref>{{cite journal | vauthors = Yang D, Singh A, Wu H, Kroe-Barrett R | title = Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics | journal = Analytical Biochemistry | volume = 508 | pages = 78–96 | date = September 2016 | pmid = 27365220 | doi = 10.1016/j.ab.2016.06.024 | doi-access = free }}</ref>

==See also==