Implement enhanced glycan sensors with control algorithms to produce consistent, desirable glycosylation patterns to improve product quality of biotherapeutics.
A bioprocess performance platform producing near real-time control of critical glycan attributes will be ready for pilot scale implementation.
Improved process efficiency and reduced batch-to-batch variability
We have developed sensor detection interfaces that can interrogate antibody titer and glycosylation, respectively, through the transduction of an electrochemical output. These interfaces make use of a novel PEG thiol-mediated assembly methodology and are rapidly electroassembled onto sensor electrodes. For titer detection interface, we have developed a cysteine-tagged protein G antibody recognition protein, that when linked the thiolated PEG interface, enables rapid, robust assessment of total antibody concentration. For glycosylation detection, instead of assembling the cysteinylated protein G, thiolated sugar groups occupy many of the open thiol-PEG binding sites so that the sugar groups act as a lectin bait. Then, sugar-specific lectins are layered on top of the thiolated PEG hydrogel. These surfaces then provide selective capture of antibodies based on the traditional lectin-glycan binding. Further, when both interfaces are coupled with an electrochemical reporter, these interfaces enable near real time electrochemical outputs.
Li, J., Maniar, D., Qu, X., Liu, H., Tsao, C., Kim, E., Bentley, W. E., Liu, C., & Payne, G. F. (2019). Coupling Self-Assembly Mechanisms to Fabricate Molecularly and Electrically Responsive Films. Biomacromolecules, 20(2), 969-978. https://doi.org/10.1021/acs.biomac.8b01592
Luo, Y., Lovelett, R. J., Price, J. V., Radhakrishnan, D., Barnthouse, K., Hu, P., Schaefer, E., Cunningham, J., Lee, K. H., Shivappa, R. B., & Ogunnaike, B. A. (2020). Modeling the Effect of Amino Acids and Copper on Monoclonal Antibody Productivity and Glycosylation: A Modular Approach. Biotechnology Journal, 16(2). https://doi.org/10.1002/biot.202000261
Mao, L., Schneider, J. W., & Robinson, A. S. (2023). Use of single analytic tool to quantify both absolute N-glycosylation and glycan distribution in monoclonal antibodies. Biotechnology Progress, 39(5). https://doi.org/10.1002/btpr.3365
Motabar, D., Li, J., Wang, S., Tsao, C., Tong, X., Wang, L., Payne, G. F., & Bentley, W. E. (2021). Simple, rapidly electroassembled thiolated PEG-based sensor interfaces enable rapid interrogation of antibody titer and glycosylation. Biotechnology and Bioengineering, 118(7), 2744-2758. https://doi.org/10.1002/bit.27793
Wells, E., Song, L., Greer, M., Luo, Y., Kurian, V., Ogunnaike, B., & Robinson, A. S. (2020). Media supplementation for targeted manipulation of monoclonal antibody galactosylation and fucosylation. Biotechnology and Bioengineering, 117(11), 3310-3321. https://doi.org/10.1002/bit.27496
Adaptive Process Control and Advanced Sensing for Robust mAb Glycan Quality (QSP1.0-023), NIIMBL Member Forum, Virtual, May 28, 2020.
Adaptive Process Control and Advanced Sensing for Robust mAb Glycan Quality (QSP1.0-023), NIIMBL Member Forum, Virtual, May 30, 2019.
Chen, S. & Zhang, H., High Throughput Intact Glycopeptide Enrichment for Site-specific N-linked Glycosylation Analysis Utilizing Automated Liquid Handling Systems, 67th ASMS Conference on Mass Spectrometry and Allied Topics, Atlanta, GA, June 2, 2019.
Luo, Y., Control of Glycosylation and Titer in Fed-Batch Monoclonal Antibody Production, ACS National Meeting, Philadelphia, PA, March 22, 2020.
Luo, Y., Control of Glycosylation and Titer in Fed-Batch Monoclonal Antibody Production, AIChE Annual Meeting, Orlando, FL, November 14, 2019.
Robinson, A. S., Controlling Protein Quality and Antibody Expression, Protein Australia, Brisbane, Australia, July 16, 2019.
Robinson, A., Controlling Protein Quality and Antibody Expression, Peptalk 2019, San Diego, CA, January 14, 2019.
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