Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance
ABSTRACTAccurate and efficient affinity measurement techniques are essential for the biophysical characterization of therapeutic monoclonal antibodies, one of the fastest growing drug classes. Surface plasmon resonance (SPR) is widely used for determining antibody affinity, but does not perform well...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2023-12-01
|
| Series: | mAbs |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2023.2291209 |
| _version_ | 1797356279043194880 |
|---|---|
| author | M. Frank Erasmus Molly Dovner Fortunato Ferrara Sara D’Angelo André A. Teixeira Camila Leal-Lopes Laura Spector Elizabeth Hopkins Andrew R. M. Bradbury |
| author_facet | M. Frank Erasmus Molly Dovner Fortunato Ferrara Sara D’Angelo André A. Teixeira Camila Leal-Lopes Laura Spector Elizabeth Hopkins Andrew R. M. Bradbury |
| author_sort | M. Frank Erasmus |
| collection | DOAJ |
| description | ABSTRACTAccurate and efficient affinity measurement techniques are essential for the biophysical characterization of therapeutic monoclonal antibodies, one of the fastest growing drug classes. Surface plasmon resonance (SPR) is widely used for determining antibody affinity, but does not perform well with extremely high affinity (low picomolar to femtomolar range) molecules. In this study, we compare the SPR-based Carterra LSA and the kinetic exclusion assay (KinExA) for measuring the affinities of 48 antibodies generated against the SARS-CoV-2 receptor-binding domain. These data reveal that high-affinity antibodies can be generated straight from selections using high-quality in vitro library platforms with 54% correspondence between affinities measured using LSA and KinExA. Generally, where there was a 2-fold or greater difference between LSA and KinExA, KinExA reported that affinities were tighter. We highlight the differences between LSA and KinExA, identifying the benefits and pitfalls of each in terms of dynamic range and throughput. Furthermore, we demonstrate for the first time that single-point screening with KinExA can significantly improve throughput while maintaining a strong correlation with full binding curve equilibrium measurements, enabling the accurate rank-ordering of clones with exceptionally tight binding properties. |
| first_indexed | 2024-03-08T14:24:19Z |
| format | Article |
| id | doaj.art-8d9d6973ecb14fbf9cc087829f04f11c |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2024-03-08T14:24:19Z |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-8d9d6973ecb14fbf9cc087829f04f11c2024-01-13T11:27:51ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2291209Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonanceM. Frank Erasmus0Molly Dovner1Fortunato Ferrara2Sara D’Angelo3André A. Teixeira4Camila Leal-Lopes5Laura Spector6Elizabeth Hopkins7Andrew R. M. Bradbury8Specifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASapidyne, Inc, Boise, ID, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USASapidyne, Inc, Boise, ID, USASpecifica, LLC, a Q2 Solutions Company, Santa Fe, NM, USAABSTRACTAccurate and efficient affinity measurement techniques are essential for the biophysical characterization of therapeutic monoclonal antibodies, one of the fastest growing drug classes. Surface plasmon resonance (SPR) is widely used for determining antibody affinity, but does not perform well with extremely high affinity (low picomolar to femtomolar range) molecules. In this study, we compare the SPR-based Carterra LSA and the kinetic exclusion assay (KinExA) for measuring the affinities of 48 antibodies generated against the SARS-CoV-2 receptor-binding domain. These data reveal that high-affinity antibodies can be generated straight from selections using high-quality in vitro library platforms with 54% correspondence between affinities measured using LSA and KinExA. Generally, where there was a 2-fold or greater difference between LSA and KinExA, KinExA reported that affinities were tighter. We highlight the differences between LSA and KinExA, identifying the benefits and pitfalls of each in terms of dynamic range and throughput. Furthermore, we demonstrate for the first time that single-point screening with KinExA can significantly improve throughput while maintaining a strong correlation with full binding curve equilibrium measurements, enabling the accurate rank-ordering of clones with exceptionally tight binding properties.https://www.tandfonline.com/doi/10.1080/19420862.2023.2291209Antibodydrug discoveryantibody therapeuticssurface plasmon resonancekinetic exclusion assayantibody characterization |
| spellingShingle | M. Frank Erasmus Molly Dovner Fortunato Ferrara Sara D’Angelo André A. Teixeira Camila Leal-Lopes Laura Spector Elizabeth Hopkins Andrew R. M. Bradbury Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance mAbs Antibody drug discovery antibody therapeutics surface plasmon resonance kinetic exclusion assay antibody characterization |
| title | Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance |
| title_full | Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance |
| title_fullStr | Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance |
| title_full_unstemmed | Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance |
| title_short | Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance |
| title_sort | determining the affinities of high affinity antibodies using kinexa and surface plasmon resonance |
| topic | Antibody drug discovery antibody therapeutics surface plasmon resonance kinetic exclusion assay antibody characterization |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2023.2291209 |
| work_keys_str_mv | AT mfrankerasmus determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT mollydovner determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT fortunatoferrara determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT saradangelo determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT andreateixeira determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT camilaleallopes determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT lauraspector determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT elizabethhopkins determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance AT andrewrmbradbury determiningtheaffinitiesofhighaffinityantibodiesusingkinexaandsurfaceplasmonresonance |