Abstract
Standard antibody phage-display panning uses purified proteins, antigen-transfected cells, or tumor cell lines as target structure to generate specific antibodies. Here, we describe a method for the selection of specific antibodies by phage panning against routine formalin-fixed paraffin-embedded (FFPE) tissue biopsies immobilized on glass slides. Selected antibody fragments recognize disease-associated antigens in its native conformation, suitable for the development of targeted diagnostic and therapeutic agents.
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References
Fleuren ED, Versleijen-Jonkers YM, Heskamp S, van Herpen CM, Oyen WJ, van der Graaf WT, Boerman OC (2014) Theranostic applications of antibodies in oncology. Mol Oncol 8(4):799–812. https://doi.org/10.1016/j.molonc.2014.03.010
Schofield DJ, Lewis AR, Austin MJ (2014) Genetic methods of antibody generation and their use in immunohistochemistry. Methods 70(1):20–27. https://doi.org/10.1016/j.ymeth.2014.02.031
Sanchez-Martin D, Sorensen MD, Lykkemark S, Sanz L, Kristensen P, Ruoslahti E, Alvarez-Vallina L (2015) Selection strategies for anticancer antibody discovery: searching off the beaten path. Trends Biotechnol 33(5):292–301. https://doi.org/10.1016/j.tibtech.2015.02.008
Ten Haaf A, Pscherer S, Fries K, Barth S, Gattenlohner S, Tur MK (2015) Phage display-based on-slide selection of tumor-specific antibodies on formalin-fixed paraffin-embedded human tissue biopsies. Immunol Lett 166(2):65–78. https://doi.org/10.1016/j.imlet.2015.05.013
Jarutat T, Nickels C, Frisch C, Stellmacher F, Hofig KP, Knappik A, Merz H (2007) Selection of vimentin-specific antibodies from the HuCAL phage display library by subtractive panning on formalin-fixed, paraffin-embedded tissue. Biol Chem 388(6):651–658. https://doi.org/10.1515/BC.2007.070
Sun Y, Shukla GS, Kennedy GG, Warshaw DM, Weaver DL, Pero SC, Floyd L, Krag DN (2009) Biopanning phage-display libraries on small tissue sections captured by laser capture microdissection. J Biotech Res 1:55–63
Tanca A, Pagnozzi D, Addis MF (2012) Setting proteins free: progresses and achievements in proteomics of formalin-fixed, paraffin-embedded tissues. Proteomics Clin Appl 6(1–2):7–21. https://doi.org/10.1002/prca.201100044
Thavarajah R, Mudimbaimannar VK, Elizabeth J, Rao UK, Ranganathan K (2012) Chemical and physical basics of routine formaldehyde fixation. J Oral Maxillofac Pathol 16(3):400–405. https://doi.org/10.4103/0973-029X.102496
Tanaka T, Ito T, Furuta M, Eguchi C, Toda H, Wakabayashi-Takai E, Kaneko K (2002) In situ phage screening. A method for identification of subnanogram tissue components in situ. J Biol Chem 277(33):30382–30387. https://doi.org/10.1074/jbc.M203547200
Acknowledgment
This work was supported by the “Verein zur Förderung der Krebsforschung in Giessen e. V.”
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ten Haaf, A., Gattenlöhner, S., Tur, M.K. (2018). Antibody Selection on FFPE Tissue Slides. 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_21
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DOI: https://doi.org/10.1007/978-1-4939-7447-4_21
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