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.
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.
Improved process efficiency and reduced batch-to-batch variability
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
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
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
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
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
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