Abstract
The majority of therapeutic antibodies, bispecific antibodies, and chimeric antigen receptor (CAR) T cells in cancer therapy are based on an antibody or antibody fragment that specifically binds a target present on the surface of a tumor cell. Suitable antigens that can be used for immunotherapy are ideally tumor-specific or tumor-associated and stably expressed on the tumor cell. The identification of new target structures to further optimize immunotherapies could be realized by comparing healthy and tumor cells using “omics” methods to select promising proteins. However, differences in post-translational modifications and structural alterations that can be present on the tumor cell surface are difficult to identify or even not accessible by these techniques. In this chapter, we describe an alternative approach to potentially identify antibodies targeting novel tumor-associated antigens (TAA) or epitopes by using cellular screening and phage display of antibody libraries. Isolated antibody fragments can be further converted into chimeric IgG or other antibody formats to investigate the anti-tumor effector functions and finally identify and characterize the respective antigen.
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References
Carter PJ, Lazar GA (2018) Next generation antibody drugs: pursuit of the “high-hanging fruit”. Nat Rev Drug Discov 17(3):197–223
Carter P, Smith L, Ryan M (2004) Identification and validation of cell surface antigens for antibody targeting in oncology. Endocr Relat Cancer 11(4):659–687
Bakker AB, van den Oudenrijn S, Bakker AQ, Feller N, van Meijer M, Bia JA et al (2004) C-type lectin-like molecule-1: a novel myeloid cell surface marker associated with acute myeloid leukemia. Cancer Res 64(22):8443–8450
Ljungars A, Mårtensson L, Mattsson J, Kovacek M, Sundberg A, Tornberg UC et al (2018) A platform for phenotypic discovery of therapeutic antibodies and targets applied on Chronic Lymphocytic Leukemia. NPJ Precis Oncol 2(1):18
Alamyar E, Duroux P, Lefranc MP, Giudicelli V (2012) IMGT((R)) tools for the nucleotide analysis of immunoglobulin (IG) and T cell receptor (TR) V-(D)-J repertoires, polymorphisms, and IG mutations: IMGT/V-QUEST and IMGT/HighV-QUEST for NGS. Methods Mol Biol 882:569–604
Bystry V, Reigl T, Krejci A, Demko M, Hanakova B, Grioni A et al (2017) ARResT/Interrogate: an interactive immunoprofiler for IG/TR NGS data. Bioinformatics 33(3):435–437
Ravn U, Didelot G, Venet S, Ng KT, Gueneau F, Rousseau F et al (2013) Deep sequencing of phage display libraries to support antibody discovery. Methods 60(1):99–110
Ravn U, Gueneau F, Baerlocher L, Osteras M, Desmurs M, Malinge P et al (2010) By-passing in vitro screening--next generation sequencing technologies applied to antibody display and in silico candidate selection. Nucleic Acids Res 38(21):e193
Ljungars A, Svensson C, Carlsson A, Birgersson E, Tornberg UC, Frendeus B et al (2019) Deep mining of complex antibody phage pools generated by cell panning enables discovery of rare antibodies binding new targets and epitopes. Front Pharmacol 10:847
Krohn S, Boje AS, Gehlert CL, Lutz S, Darzentas N, Knecht H et al (2022) Identification of new antibodies targeting malignant plasma cells for immunotherapy by next-generation sequencing-assisted phage display. Front Immunol 13:908093
Burmester J, Plückthun A (2001) Construction of scFv fragments from hybridoma or spleen cells by PCR assembly. In: Kontermann R, Dübel S (eds) Antibody engineering. Springer, Berlin, Heidelberg, pp 19–40
Kay BK, Winter J, McCafferty J (1996) Phage display of peptides and proteins: a laboratory manual. Elsevier Science
Acknowledgments
The pJB12 vector was kindly provided by Professor Andreas Plückthun. This work was supported by a research grant from the Else Kröner-Fresenius-Stiftung to K.K. and a research grant from the Deutsche Krebshilfe e.V. to M.P.
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Krohn, S., Peipp, M., Klausz, K. (2023). Identification of New Antibodies Targeting Tumor Cell Surface Antigens by Phage Display. In: Zielonka, S., Krah, S. (eds) Genotype Phenotype Coupling. Methods in Molecular Biology, vol 2681. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3279-6_5
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DOI: https://doi.org/10.1007/978-1-0716-3279-6_5
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