Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor
Anthrax lethal factor (LF) is one of the enzymatic components of the anthrax toxin responsible for the pathogenic responses of the anthrax disease. The ability to screen multiplexed ligands against LF and subsequently estimate the effective kinetic rates (<inline-formula><math xmlns="h...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-11-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/11/12/483 |
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| author | Allison M. Marn James Needham Elisa Chiodi M. Selim Ünlü |
| author_facet | Allison M. Marn James Needham Elisa Chiodi M. Selim Ünlü |
| author_sort | Allison M. Marn |
| collection | DOAJ |
| description | Anthrax lethal factor (LF) is one of the enzymatic components of the anthrax toxin responsible for the pathogenic responses of the anthrax disease. The ability to screen multiplexed ligands against LF and subsequently estimate the effective kinetic rates (<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msub><mi>k</mi><mrow><mi>o</mi><mi>n</mi></mrow></msub></semantics></math></inline-formula> and <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msub><mi>k</mi><mrow><mi>o</mi><mi>f</mi><mi>f</mi></mrow></msub></semantics></math></inline-formula>) and complementary binding behavior provides critical information useful in diagnostic and therapeutic development for anthrax. Tools such as biolayer interferometry (BLI) and surface plasmon resonance imaging (SPRi) have been developed for this purpose; however, these tools suffer from limitations such as signal jumps when the solution in the chamber is switched or low sensitivity. Here, we present multiplexed antibody affinity measurements obtained by the interferometric reflectance imaging sensor (IRIS), a highly sensitive, label-free optical biosensor, whose stability, simplicity, and imaging modality overcomes many of the limitations of other multiplexed methods. We compare the multiplexed binding results obtained with the IRIS system using two ligands targeting the anthrax lethal factor (LF) against previously published results obtained with more traditional surface plasmon resonance (SPR), which showed consistent results, as well as kinetic information previously unattainable with SPR. Additional exemplary data demonstrating multiplexed binding and the corresponding complementary binding to sequentially injected ligands provides an additional layer of information immediately useful to the researcher. |
| first_indexed | 2024-03-10T04:32:50Z |
| format | Article |
| id | doaj.art-7e549b8d6388427ca1a785a9630a60fd |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-10T04:32:50Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-7e549b8d6388427ca1a785a9630a60fd2023-11-23T04:01:32ZengMDPI AGBiosensors2079-63742021-11-01111248310.3390/bios11120483Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging SensorAllison M. Marn0James Needham1Elisa Chiodi2M. Selim Ünlü3School of Engineering, Computing, and Construction Management, Roger Williams University, Bristol, RI 02809, USAInBios International, Inc., Seattle, WA 98109, USADepartment of Electrical Engineering, Boston University, Boston, MA 02215, USADepartment of Electrical Engineering, Boston University, Boston, MA 02215, USAAnthrax lethal factor (LF) is one of the enzymatic components of the anthrax toxin responsible for the pathogenic responses of the anthrax disease. The ability to screen multiplexed ligands against LF and subsequently estimate the effective kinetic rates (<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msub><mi>k</mi><mrow><mi>o</mi><mi>n</mi></mrow></msub></semantics></math></inline-formula> and <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msub><mi>k</mi><mrow><mi>o</mi><mi>f</mi><mi>f</mi></mrow></msub></semantics></math></inline-formula>) and complementary binding behavior provides critical information useful in diagnostic and therapeutic development for anthrax. Tools such as biolayer interferometry (BLI) and surface plasmon resonance imaging (SPRi) have been developed for this purpose; however, these tools suffer from limitations such as signal jumps when the solution in the chamber is switched or low sensitivity. Here, we present multiplexed antibody affinity measurements obtained by the interferometric reflectance imaging sensor (IRIS), a highly sensitive, label-free optical biosensor, whose stability, simplicity, and imaging modality overcomes many of the limitations of other multiplexed methods. We compare the multiplexed binding results obtained with the IRIS system using two ligands targeting the anthrax lethal factor (LF) against previously published results obtained with more traditional surface plasmon resonance (SPR), which showed consistent results, as well as kinetic information previously unattainable with SPR. Additional exemplary data demonstrating multiplexed binding and the corresponding complementary binding to sequentially injected ligands provides an additional layer of information immediately useful to the researcher.https://www.mdpi.com/2079-6374/11/12/483multiplexed antibody kineticsIRISinterferometric sensinganthrax lethal factor |
| spellingShingle | Allison M. Marn James Needham Elisa Chiodi M. Selim Ünlü Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor Biosensors multiplexed antibody kinetics IRIS interferometric sensing anthrax lethal factor |
| title | Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor |
| title_full | Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor |
| title_fullStr | Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor |
| title_full_unstemmed | Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor |
| title_short | Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor |
| title_sort | multiplexed high sensitivity measurements of antibody affinity using interferometric reflectance imaging sensor |
| topic | multiplexed antibody kinetics IRIS interferometric sensing anthrax lethal factor |
| url | https://www.mdpi.com/2079-6374/11/12/483 |
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