Abstract
The application of recombinant human antibodies is growing rapidly mainly in the field of diagnostics and therapeutics. To identify antibodies against a specific antigen, panning selection is carried out using different display technologies. Phage display technology remains the preferred platform due to its robustness and efficiency in biopanning experiments. There are both manual and semi-automated panning selections using polystyrene plastic, magnetic beads, and nitrocellulose as the immobilizing solid surface. Magnetic nanoparticles allow for improved antigen binding due to their large surface area. The Kingfisher Flex magnetic particle processing system was originally designed to aid in RNA, DNA, and protein extraction using magnetic beads. However, the system can be programmed for antibody phage display panning. The automation allows for a reduction in human error and improves reproducibility in between selections with the preprogrammed movements. The system requires minimum human intervention to operate; however, human intervention is needed for post-panning steps like phage rescue. In addition, polyclonal and monoclonal ELISA can be performed using the semi-automated platform to evaluate the selected antibody clones. This chapter will summarize the suggested protocol from the panning stage till the monoclonal ELISA evaluation. Other than this, important notes on the possible optimization and troubleshooting are also included at the end of this chapter.
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Acknowledgment
The authors would like to acknowledge the support of USM Research University Individual grant (1001/CIPPM/812173) and Malaysian Ministry of Education through the Higher Institution Centre of Excellence (HICoE) Grant (Grant No: 311/CIPPM/44001005).
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Ch’ng, A.C.W., Hamidon, N.H.B., Konthur, Z., Lim, T.S. (2018). Magnetic Nanoparticle-Based Semi-Automated Panning for High-Throughput Antibody Selection. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_16
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DOI: https://doi.org/10.1007/978-1-4939-7447-4_16
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