Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics
Open accessAbstract
The acquisition of reliable kinetic parameters for the characterization of biomolecular interactions is an important component of the drug discovery and development process. While several benchmark studies have explored the variability of kinetic rate constants obtained from multiple laboratories and biosensors, a direct comparison of these instruments' performance has not been undertaken, and systematic factors contributing to data variability from these systems have not been discussed. To address these questions, a panel of ten high-affinity monoclonal antibodies was simultaneously evaluated for their binding kinetics against the same antigen on four biosensor platforms: GE Healthcare's Biacore T100, Bio-Rad's ProteOn XPR36, ForteBio's Octet RED384, and Wasatch Microfluidics's IBIS MX96. We compared the strengths and weaknesses of these systems and found that despite certain inherent systematic limitations in instrumentation, the rank orders of both the association and dissociation rate constants were highly correlated between these instruments. Our results also revealed a trade-off between data reliability and sample throughput. Biacore T100, followed by ProteOn XPR36, exhibited excellent data quality and consistency, whereas Octet RED384 and IBIS MX96 demonstrated high flexibility and throughput with compromises in data accuracy and reproducibility. Our results support the need for a "fit-for-purpose" approach in instrument selection for biosensor studies.
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Surface plasmon resonance quantifies adenoviruses in downstream samples, with low throughput being its main drawback compared to BLI.
“First, surface plasmon resonance, like BLI, quantifies the concentration of an analyte by measuring changes to the optical properties of a sensor chip, in this case, a shift in the resonance angle of free electrons exited by polarized light, caused by analyte binding to immobilized antibodies on its surface. It successfully quantifies adenoviruses in downstream samples,120 with low throughput”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [121]
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“SPR and BLI suffer from several well-established limitations, including mass transport and sensitivity to bulk shifts caused by buffer formulation that can obscure the true dynamics of the analyte–ligand interaction.(27,28) Consequently, despite robustness across diverse spot densities, antibodies, and FcγR, we were u…”
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A Systematic Approach to HIV-1 Vaccine Immunogen Selection
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“Multiple studies including a benchmark study have been conducted to compare binding affinities derived using some of these technologies [34,35].”
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“The midpoint of ELISA binding curve was used to estimate the affinity as biosensor values are unreliable for very high affinity mAbs [18].”
Performance qualification for reproducible Surface Plasmon Resonance analysis
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Detecting ligand interactions in real time on living bacterial cells
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“Due to the technical limitations of BLI measurements for high-affinity interactions (35) and the complex valency of this specific interaction, these values are considered observed KD values to guide future experiments.”
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“This approach enables real-time measurement of molecular binding kinetics, similar in principle to widely used lab equipment such as surface plasmon resonance (SPR) or biolayer interference (BLI) 8 , but with unique size and cost advantages.”
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Characterization of Critical Quality Attributes of an Anti-PCSK9 Monoclonal Antibody
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- Authors
- Yang, D., Singh, A., Wu, H., & Kroe-Barrett, R.
- Affiliations
- Boehringer Ingelheim (United States) (US)
- Venue
- Analytical Biochemistry
- Publisher
- Elsevier BV
- Published
- 2016-09
- Volume
- 508
- Pages
- 78-96
- ISSN
- 0003-2697 (ISSN-L)
- OpenAlex
- W2470598491
- Semantic Scholar
- 6448aac56c434e3377d3fd7e949f680d8bed074f
- Cited by
- 120
- References
- 24
- Citations by year
- 2026: 4 · 2025: 9 · 2024: 14 · 2023: 15 · 2022: 11 · 2021: 17 · 2020: 9 · 2019: 16 · 2018: 11 · 2017: 13 · 2016: 1
- FWCI
- 7.10
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Analytical Biochemistry (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › Biosimilars and Bioanalytical Methods
- Keywords
- antibody-antigen binding kinetics, association rate constant, dissociation rate constant, data reproducibility, sample throughput, instrument selection, biomolecular interactions
- MeSH terms
- Antibodies, Monoclonal, Antibody Affinity, Kinetics, Ligands, Protein Binding, Reproducibility of Results, Biosensing Techniques
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