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Phage Display pp 331–347Cite as

Combine Phage Antibody Display Library Selection on Patient Tissue Specimens with Laser Capture Microdissection to Identify Novel Human Antibodies Targeting Clinically Relevant Tumor Antigens

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

A functional approach to generate tumor-targeting human monoclonal antibodies is through selection of phage antibody display libraries directly on tumor cells. Although technically convenient, the use of cancer cell lines for the selection has limitations as those cell lines often undergo genetic and epigenetic changes during prolonged in vitro culture and alter their cell surface antigen expression profile. The key is to develop a technology that allows selection of phage antibody display libraries on tumor cells in situ residing in their natural tissue microenvironment. Laser capture microdissection (LCM) permits the precise procurement of tumor cells from human cancer patient tissue sections. Here, we describe a LCM-based method for selecting phage antibodies against tumor cells in situ using both fresh frozen and paraffin-embedded tissues. To restrict the selection to antibodies that bind internalizing epitopes, the method utilizes a polyclonal phage population pre-enriched for internalizing phage antibodies. The ability to recognize tumor cells in situ residing in their natural tissue microenvironment and to deliver payload intracellularly makes these LCM-selected antibodies attractive candidates for the development of targeted cancer therapeutics.

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Acknowledgment

Work in our laboratory is supported by grants from the National Institutes of Health/National Cancer Institute (R01 CA171315, R01 CA118919, and R01 CA129491). NKL received fellowship support from Basic Science Research Program of the National Research Foundation of Korea (NRF) that is funded by the Ministry of Education, Science and Technology (2013R1A6A3A03060495).

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Authors and Affiliations

  1. Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, 1001 Potrero Ave., 1305, San Francisco, CA, 94110, USA

    Yang Su, Scott Bidlingmaier, Nam-Kyung Lee & Bin Liu

Authors
  1. Yang Su
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  2. Scott Bidlingmaier
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  3. Nam-Kyung Lee
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  4. Bin Liu
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Corresponding author

Correspondence to Bin Liu .

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Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Su, Y., Bidlingmaier, S., Lee, NK., Liu, B. (2018). Combine Phage Antibody Display Library Selection on Patient Tissue Specimens with Laser Capture Microdissection to Identify Novel Human Antibodies Targeting Clinically Relevant Tumor Antigens. 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_18

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

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