A novel, label-free, pre-equilibrium assay to determine the association and dissociation rate constants of therapeutic antibodies on living cells.
affinity
antibody
binding kinetics
cell-based assay
label free
pre-equilibrium
rate constants
Journal
British journal of pharmacology
ISSN: 1476-5381
Titre abrégé: Br J Pharmacol
Pays: England
ID NLM: 7502536
Informations de publication
Date de publication:
02 Oct 2023
02 Oct 2023
Historique:
revised:
19
09
2023
received:
15
03
2023
accepted:
23
09
2023
pubmed:
3
10
2023
medline:
3
10
2023
entrez:
2
10
2023
Statut:
aheadofprint
Résumé
Monoclonal antibodies (Ab) represent the fastest growing drug class. Knowledge of the biophysical parameters (k We developed a pre-equilibrium kinetic exclusion assay using recent mathematical advances to determine the k Using our novel assay, we demonstrated for several monoclonal Ab:receptor pairs that the calculated kinetic rate constants were comparable with orthogonal methods that were lower throughput or more resource consuming. We ran simulations to predict the critical conditions to improve the performance of the assays. We further showed that this method could successfully be applied to both suspension and adherent cells. Finally, we demonstrated that k Our novel assay has the potential to systematically probe binding kinetics of monoclonal Abs to cells and can be incorporated in a screening cascade to identify new therapeutic candidates. Wide-spread adoption of pre-equilibrium assays using physiologically relevant systems will lead to a more holistic understanding of how Ab binding kinetics influence their potency.
Sections du résumé
BACKGROUND AND PURPOSE
OBJECTIVE
Monoclonal antibodies (Ab) represent the fastest growing drug class. Knowledge of the biophysical parameters (k
EXPERIMENTAL APPROACH
METHODS
We developed a pre-equilibrium kinetic exclusion assay using recent mathematical advances to determine the k
KEY RESULTS
RESULTS
Using our novel assay, we demonstrated for several monoclonal Ab:receptor pairs that the calculated kinetic rate constants were comparable with orthogonal methods that were lower throughput or more resource consuming. We ran simulations to predict the critical conditions to improve the performance of the assays. We further showed that this method could successfully be applied to both suspension and adherent cells. Finally, we demonstrated that k
CONCLUSIONS AND IMPLICATIONS
CONCLUSIONS
Our novel assay has the potential to systematically probe binding kinetics of monoclonal Abs to cells and can be incorporated in a screening cascade to identify new therapeutic candidates. Wide-spread adoption of pre-equilibrium assays using physiologically relevant systems will lead to a more holistic understanding of how Ab binding kinetics influence their potency.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Genentech, Inc
Informations de copyright
© 2023 British Pharmacological Society.
Références
Agnolon, V., Contato, A., Meneghello, A., Tagliabue, E., Toffoli, G., Gion, M., Polo, F., & Fabricio, A. S. C. (2020). ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab. Scientific Reports, 10(1), 3016. https://doi.org/10.1038/s41598-020-59630-y
Alexander, S. P., Kelly, E., Mathie, A., Peters, J. A., Veale, E. L., Armstrong, J. F., Faccenda, E., Harding, S. D., Pawson, A. J., Southan, C., Buneman, O. P., Cidlowski, J. A., Christopoulos, A., Davenport, A. P., Fabbro, D., Spedding, M., Striessnig, J., Davies, J. A., Ahlers-Dannen, K. E., … Zolghadri, Y. (2021). THE CONCISE GUIDE TO PHARMACOLOGY 2021/22: Introduction and other protein targets. British Journal of Pharmacology and Chemotherapy, 178(Suppl 1), S1-S26. https://doi.org/10.1111/bph.15537
Andersson, K., Björkelund, H., & Malmqvist, M. (2010). Antibody-antigen interactions: What is the required time to equilibrium? Nature Precedings. https://doi.org/10.1038/npre.2010.5218.1
Andersson, K., Karlsson, R., Lofas, S., Franklin, G., & Hamalainen, M. D. (2006). Label-free kinetic binding data as a decisive element in drug discovery. Expert Opinion on Drug Discovery, 1(5), 439-446. https://doi.org/10.1517/17460441.1.5.439
Bjorke, H., & Andersson, K. (2006). Measuring the affinity of a radioligand with its receptor using a rotating cell dish with in situ reference area. Applied Radiation and Isotopes, 64(1), 32-37. https://doi.org/10.1016/j.apradiso.2005.06.007
Bondza, S., Bjorkelund, H., Nestor, M., Andersson, K., & Buijs, J. (2017). Novel real-time proximity assay for characterizing multiple receptor interactions on living cells. Analytical Chemistry, 89(24), 13212-13218. https://doi.org/10.1021/acs.analchem.7b02983
Bondza, S., Foy, E., Brooks, J., Andersson, K., Robinson, J., Richalet, P., & Buijs, J. (2017). Real-time characterization of antibody binding to receptors on living immune cells. Frontiers in Immunology, 8, 455. https://doi.org/10.3389/fimmu.2017.00455
Bondza, S., Ten Broeke, T., Nestor, M., Leusen, J. H. W., & Buijs, J. (2020). Bivalent binding on cells varies between anti-CD20 antibodies and is dose-dependent. MAbs, 12(1), 1792673. https://doi.org/10.1080/19420862.2020.1792673
Bravo, D. D., Shi, Y., Sheu, A., Liang, W. C., Lin, W., Wu, Y., Yan, M., & Wang, J. (2023). A real-time image-based Efferocytosis assay for the discovery of functionally inhibitory anti-MerTK antibodies. Journal of Immunology, 210, 1166-1176. https://doi.org/10.4049/jimmunol.2200597
Brinkmann, U., & Kontermann, R. E. (2017). The making of bispecific antibodies. MAbs, 9(2), 182-212. https://doi.org/10.1080/19420862.2016.1268307
Carter, P. J., & Lazar, G. A. (2018). Next generation antibody drugs: Pursuit of the ‘high-hanging fruit’. Nature Reviews. Drug Discovery, 17(3), 197-223. https://doi.org/10.1038/nrd.2017.227
Chodorge, M., Zuger, S., Stirnimann, C., Briand, C., Jermutus, L., Grutter, M. G., & Minter, R. R. (2012). A series of Fas receptor agonist antibodies that demonstrate an inverse correlation between affinity and potency. Cell Death and Differentiation, 19(7), 1187-1195. https://doi.org/10.1038/cdd.2011.208
Chung, I., Reichelt, M., Shao, L., Akita, R. W., Koeppen, H., Rangell, L., Schaefer, G., Mellman, I., & Sliwkowski, M. X. (2016). High cell-surface density of HER2 deforms cell membranes. Nature Communications, 7, 12742. https://doi.org/10.1038/ncomms12742
Cooper, M. A. (2002). Optical biosensors in drug discovery. Nature Reviews. Drug Discovery, 1(7), 515-528. https://doi.org/10.1038/nrd838
Copeland, R. A. (2016). The drug-target residence time model: A 10-year retrospective. Nature Reviews. Drug Discovery, 15(2), 87-95. https://doi.org/10.1038/nrd.2015.18
Copeland, R. A., Pompliano, D. L., & Meek, T. D. (2006). Drug-target residence time and its implications for lead optimization. Nature Reviews. Drug Discovery, 5(9), 730-739. https://doi.org/10.1038/nrd2082
Curtis, M. J., Alexander, S. P. H., Cirino, G., George, C. H., Kendall, D. A., Insel, P. A., Izzo, A. A., Ji, Y., Panettieri, R. A., Patel, H. H., Sobey, C. G., Stanford, S. C., Stanley, P., Stefanska, B., Stephens, G. J., Teixeira, M. M., Vergnolle, N., & Ahluwalia, A. (2022). Planning experiments: Updated guidance on experimental design and analysis and their reporting III. British Journal of Pharmacology, 179(15), 3907-3913. https://doi.org/10.1111/bph.15868
Dahl, G., & Akerud, T. (2013). Pharmacokinetics and the drug-target residence time concept. Drug Discovery Today, 18(15-16), 697-707. https://doi.org/10.1016/j.drudis.2013.02.010
Dai, Z., Juneja, J., Schneeweis, L., Cohen, D., Marsilio, F., Morin, P., & DasGupta, R. (2020). Application of the Gyrolab microfluidic platform to measure picomolar affinity of a PD-L1-binding Adnectin™ radioligand for positron emission tomography. BioTechniques, 69(3), 200-205. https://doi.org/10.2144/btn-2020-0080
Day, E. S., Capili, A. D., Borysenko, C. W., Zafari, M., & Whitty, A. (2013). Determining the affinity and stoichiometry of interactions between unmodified proteins in solution using Biacore. Analytical Biochemistry, 440(1), 96-107. https://doi.org/10.1016/j.ab.2013.05.012
Dela Cruz Chuh, J., Go, M., Chen, Y., Guo, J., Rafidi, H., Mandikian, D., Sun, Y., Lin, Z., Schneider, K., Zhang, P., Vij, R., Sharpnack, D., Chan, P., de la Cruz, C., Sadowsky, J., Seshasayee, D., Koerber, J. T., Pillow, T. H., Phillips, G. D., … Agard, N. J. (2021). Preclinical optimization of Ly6E-targeted ADCs for increased durability and efficacy of anti-tumor response. MAbs, 13(1), 1862452. https://doi.org/10.1080/19420862.2020.1862452
Dong, T., Han, C., Liu, X., Wang, Z., Wang, Y., Kang, Q., Wang, P., & Zhou, F. (2023). Live cells versus fixated cells: Kinetic measurements of biomolecular interactions with the LigandTracer method and surface plasmon resonance microscopy. Molecular Pharmaceutics, 20(4), 2094-2104. https://doi.org/10.1021/acs.molpharmaceut.2c01047
Drake, A. W., Myszka, D. G., & Klakamp, S. L. (2004). Characterizing high-affinity antigen/antibody complexes by kinetic- and equilibrium-based methods. Analytical Biochemistry, 328(1), 35-43. https://doi.org/10.1016/j.ab.2003.12.025
Drake, A. W., Tang, M. L., Papalia, G. A., Landes, G., Haak-Frendscho, M., & Klakamp, S. L. (2012). Biacore surface matrix effects on the binding kinetics and affinity of an antigen/antibody complex. Analytical Biochemistry, 429(1), 58-69. https://doi.org/10.1016/j.ab.2012.06.024
Engelberts, P. J., Voorhorst, M., Schuurman, J., van Meerten, T., Bakker, J. M., Vink, T., Mackus, W. J., Breij, E. C., Derer, S., Valerius, T., van de Winkel, J. G., Parren, P. W., & Beurskens, F. J. (2016). Type I CD20 antibodies recruit the B cell receptor for complement-dependent lysis of malignant B cells. Journal of Immunology, 197(12), 4829-4837. https://doi.org/10.4049/jimmunol.1600811
Estep, P., Reid, F., Nauman, C., Liu, Y., Sun, T., Sun, J., & Xu, Y. (2013). High throughput solution-based measurement of antibody-antigen affinity and epitope binning. MAbs, 5(2), 270-278. https://doi.org/10.4161/mabs.23049
Hadzhieva, M., Pashov, A. D., Kaveri, S., Lacroix-Desmazes, S., Mouquet, H., & Dimitrov, J. D. (2017). Impact of antigen density on the binding mechanism of IgG antibodies. Scientific Reports, 7(1), 3767. https://doi.org/10.1038/s41598-017-03942-z
Hein, P., Michel, M. C., Leineweber, K., Wieland, T., Wettschureck, N., & Offermanns, S. (2005). Receptor and binding studies. In S. Dhein, F. W. Mohr, & M. Delmar (Eds.), Practical methods in cardiovascular research (pp. 723-783). Springer. https://doi.org/10.1007/3-540-26574-0_37
Hendriks, B. S., Klinz, S. G., Reynolds, J. G., Espelin, C. W., Gaddy, D. F., & Wickham, T. J. (2013). Impact of tumor HER2/ERBB2 expression level on HER2-targeted liposomal doxorubicin-mediated drug delivery: Multiple low-affinity interactions lead to a threshold effect. Molecular Cancer Therapeutics, 12(9), 1816-1828. https://doi.org/10.1158/1535-7163.MCT-13-0180
Hunter, S. A., & Cochran, J. R. (2016). Cell-binding assays for determining the affinity of protein-protein interactions: Technologies and considerations. Methods in Enzymology, 580, 21-44. https://doi.org/10.1016/bs.mie.2016.05.002
Junttila, T. T., Akita, R. W., Parsons, K., Fields, C., Lewis Phillips, G. D., Friedman, L. S., Sampath, D., & Sliwkowski, M. X. (2009). Ligand-independent HER2/HER3/PI3K complex is disrupted by trastuzumab and is effectively inhibited by the PI3K inhibitor GDC-0941. Cancer Cell, 15(5), 429-440. https://doi.org/10.1016/j.ccr.2009.03.020
Karlsson, R. (1994). Real-time competitive kinetic analysis of interactions between low-molecular-weight ligands in solution and surface-immobilized receptors. Analytical Biochemistry, 221(1), 142-151. https://doi.org/10.1006/abio.1994.1390
Kielczewska, A., D'Angelo, I., Amador, M. S., Wang, T., Sudom, A., Min, X., Rathanaswami, P., Pigott, C., & Foltz, I. N. (2022). Development of a potent high-affinity human therapeutic antibody via novel application of recombination signal sequence-based affinity maturation. The Journal of Biological Chemistry, 298(2), 101533. https://doi.org/10.1016/j.jbc.2021.101533
Kim, P., Liu, X., Lee, T., Liu, L., Barham, R., Kirkland, R., Leesman, G., Kuller, A., Ybarrondo, B., Ng, S. C., & Singh, S. (2011). Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients. Proteome Science, 9(1), 75. https://doi.org/10.1186/1477-5956-9-75
Leung, K. M., Batey, S., Rowlands, R., Isaac, S. J., Jones, P., Drewett, V., Carvalho, J., Gaspar, M., Weller, S., Medcalf, M., Wydro, M. M., Pegram, R., Mudde, G. C., Bauer, A., Moulder, K., Woisetschlager, M., Tuna, M., Haurum, J. S., & Sun, H. (2015). A HER2-specific modified Fc fragment (Fcab) induces antitumor effects through degradation of HER2 and apoptosis. Molecular Therapy, 23(11), 1722-1733. https://doi.org/10.1038/mt.2015.127
Maass, K. F., Kulkarni, C., Betts, A. M., & Wittrup, K. D. (2016). Determination of cellular processing rates for a trastuzumab-maytansinoid antibody-drug conjugate (ADC) highlights key parameters for ADC design. The AAPS Journal, 18(3), 635-646. https://doi.org/10.1208/s12248-016-9892-3
Natsume, A., Shimizu-Yokoyama, Y., Satoh, M., Shitara, K., & Niwa, R. (2009). Engineered anti-CD20 antibodies with enhanced complement-activating capacity mediate potent anti-lymphoma activity. Cancer Science, 100(12), 2411-2418. https://doi.org/10.1111/j.1349-7006.2009.01327.x
Noppen, B., Vanbelle, A., Stitt, A. W., & Vanhove, M. (2021). A novel assay based on pre-equilibrium titration curves for the determination of enzyme inhibitor binding kinetics. European Biophysics Journal, 50(7), 1037-1043. https://doi.org/10.1007/s00249-021-01554-0
Ohmura, N., Lackie, S. J., & Saiki, H. (2001). An immunoassay for small analytes with theoretical detection limits. Analytical Chemistry, 73(14), 3392-3399. https://doi.org/10.1021/ac001328d
Rathanaswami, P., Babcook, J., & Gallo, M. (2008). High-affinity binding measurements of antibodies to cell-surface-expressed antigens. Analytical Biochemistry, 373(1), 52-60. https://doi.org/10.1016/j.ab.2007.08.014
Salimi-Moosavi, H., Rathanaswami, P., Rajendran, S., Toupikov, M., & Hill, J. (2012). Rapid affinity measurement of protein-protein interactions in a microfluidic platform. Analytical Biochemistry, 426(2), 134-141. https://doi.org/10.1016/j.ab.2012.04.023
Sharma, S., Li, Z., Bussing, D., & Shah, D. K. (2020). Evaluation of quantitative relationship between target expression and antibody-drug conjugate exposure inside cancer cells. Drug Metabolism and Disposition, 48(5), 368-377. https://doi.org/10.1124/dmd.119.089276
Singh, V., Gupta, D., Arora, R., Tripathi, R. P., Almasan, A., & Macklis, R. M. (2014). Surface levels of CD20 determine anti-CD20 antibodies mediated cell death in vitro. PLoS ONE, 9(11), e111113. https://doi.org/10.1371/journal.pone.0111113
Slaga, D., Ellerman, D., Lombana, T. N., Vij, R., Li, J., Hristopoulos, M., Clark, R., Johnston, J., Shelton, A., Mai, E., Gadkar, K., Lo, A. A., Koerber, J. T., Totpal, K., Prell, R., Lee, G., Spiess, C., & Junttila, T. T. (2018). Avidity-based binding to HER2 results in selective killing of HER2-overexpressing cells by anti-HER2/CD3. Science Translational Medicine, 10(463), eaat5775. https://doi.org/10.1126/scitranslmed.aat5775
Spiegelberg, D., Stenberg, J., Richalet, P., & Vanhove, M. (2021). KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements. European Biophysics Journal, 50(7), 979-991. https://doi.org/10.1007/s00249-021-01560-2
Troise, F., Cafaro, V., Giancola, C., D'Alessio, G., & De Lorenzo, C. (2008). Differential binding of human immunoagents and Herceptin to the ErbB2 receptor. The FEBS Journal, 275(20), 4967-4979. https://doi.org/10.1111/j.1742-4658.2008.06625.x
Vaish, A., Lin, J. S., McBride, H. J., Grandsard, P. J., & Chen, Q. (2020). Binding affinity determination of therapeutic antibodies to membrane protein targets: Kinetic Exclusion Assay using cellular membranes for anti-CD20 antibody. Analytical Biochemistry, 609, 113974. https://doi.org/10.1016/j.ab.2020.113974
Vanhove, E., & Vanhove, M. (2018). Affinity determination of biomolecules: A kinetic model for the analysis of pre-equilibrium titration curves. European Biophysics Journal, 47(8), 961-966. https://doi.org/10.1007/s00249-018-1318-y
Vanhove, M. (2021). A method to assess the robustness of complex mathematical models used for quantitative interpretation of experimental data by nonlinear regression analysis: Application to high-affinity binding models. European Biophysics Journal, 50(7), 1045-1054. https://doi.org/10.1007/s00249-021-01556-y
Wang, X., Phan, M. M., Li, J., Gill, H., Williams, S., Gupta, N., Quarmby, V., & Yang, J. (2020). Molecular interaction characterization strategies for the development of new biotherapeutic antibody modalities. Antibodies (Basel), 9(2), 7. https://doi.org/10.3390/antib9020007
Xie, L., Mark Jones, R., Glass, T. R., Navoa, R., Wang, Y., & Grace, M. J. (2005). Measurement of the functional affinity constant of a monoclonal antibody for cell surface receptors using kinetic exclusion fluorescence immunoassay. Journal of Immunological Methods, 304(1-2), 1-14. https://doi.org/10.1016/j.jim.2005.04.009
Xie, V. (2022). Effective biomarker measurement is key for biotherapeutic development. Bioanalysis, 14(8), 451-453. https://doi.org/10.4155/bio-2022-0062
Yang, D., Singh, A., Wu, H., & Kroe-Barrett, R. (2016). Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics. Analytical Biochemistry, 508, 78-96. https://doi.org/10.1016/j.ab.2016.06.024
Zhang, N., Liu, L., Dan Dumitru, C., Cummings, N. R., Cukan, M., Jiang, Y., Li, Y., Li, F., Mitchell, T., Mallem, M. R., Ou, Y., Patel, R. N., Vo, K., Wang, H., Burnina, I., Choi, B. K., Huber, H. E., Stadheim, T. A., & Zha, D. (2011). Glycoengineered Pichia produced anti-HER2 is comparable to trastuzumab in preclinical study. MAbs, 3(3), 289-298. https://doi.org/10.4161/mabs.3.3.15532
Zhou, Y., Fei, M., Zhang, G., Liang, W. C., Lin, W., Wu, Y., Piskol, R., Ridgway, J., McNamara, E., Huang, H., Zhang, J., Oh, J., Patel, J. M., Jakubiak, D., Lau, J., Blackwood, B., Bravo, D. D., Shi, Y., Wang, J., … Yan, M. (2020). Blockade of the phagocytic receptor MerTK on tumor-associated macrophages enhances P2X7R-dependent STING activation by tumor-derived cGAMP. Immunity, 52(2), 357-373.e9. https://doi.org/10.1016/j.immuni.2020.01.014
Zhou, Y., Goenaga, A. L., Harms, B. D., Zou, H., Lou, J., Conrad, F., Adams, G. P., Schoeberl, B., Nielsen, U. B., & Marks, J. D. (2012). Impact of intrinsic affinity on functional binding and biological activity of EGFR antibodies. Molecular Cancer Therapeutics, 11(7), 1467-1476. https://doi.org/10.1158/1535-7163.MCT-11-1038