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Updated: Jun 8, 2026
Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody
Published on: May 16, 2020
Internalizing cancer antibodies from phage libraries selected on tumor cells and yeast-displayed tumor antigens
Yu Zhou1, Hao Zou, Shaoyi Zhang
1Department of Anesthesia and Pharmaceutical Chemistry, University of California, San Francisco, Room 3C-38,San Francisco General Hospital, 1001 Potrero Avenue, San Francisco, CA 94110, USA.
Abstract:
A number of approaches have been utilized to generate antibodies to cancer cell surface receptors that can be used as potential therapeutics. A number of these therapeutic approaches, including antibody-drug conjugates, immunotoxins, and targeted nucleic acid delivery, require antibodies that not only bind receptor but also undergo internalization into the cell upon binding. We previously reported on the ability to generate cancer cell binding and internalizing antibodies directly from human phage antibody libraries selected for internalization into cancer cell lines. While a number of useful antibodies have been generated using this approach, limitations include the inability to direct the selections to specific antigens and to identify the antigen bound by the antibodies. Here we show that these limitations can be overcome by using yeast-displayed antigens known to be associated with a cell type to select the phage antibody output after several rounds of selection on a mammalian cell line. We used this approach to generate several human phage antibodies to yeast-displayed EphA2 and CD44. The antibodies bound both yeast-displayed and mammalian cell surface antigens, and were endocytosed upon binding to mammalian cells. This approach is generalizable to many mammalian cell surface proteins, results in the generation of functional internalizing antibodies, and does not require antigen expression and purification for antibody generation.
Insights
Researchers developed a new method to create cancer-targeting antibodies that enter cells. This technique uses yeast-displayed antigens to improve antibody specificity and function for therapeutic applications.
Area of Science:
- Immunology
- Biotechnology
- Oncology
Background:
- Therapeutic antibodies targeting cancer cell surface receptors are crucial for treatments like antibody-drug conjugates.
- Antibodies must bind and internalize into cancer cells for efficacy in targeted therapies.
- Previous methods generated internalizing antibodies but lacked antigen specificity and identification.
Purpose of the Study:
- To overcome limitations in generating antigen-specific, internalizing antibodies for cancer therapeutics.
- To develop a method for selecting antibodies against specific cell surface proteins without prior purification.
Main Methods:
- Utilized yeast-displayed antigens (EphA2, CD44) to select phage antibody output.
- Performed selection rounds on mammalian cell lines after initial yeast display screening.
- Generated human phage antibodies against specific cell surface targets.
Main Results:
- Developed human phage antibodies that bind both yeast-displayed and mammalian cell surface antigens.
- Confirmed that generated antibodies bind to EphA2 and CD44 on mammalian cells.
- Demonstrated antibody-mediated endocytosis upon binding to target cells.
- Showcased the generalizability of the approach to various mammalian cell surface proteins.
Conclusions:
- The novel approach successfully generates functional, internalizing antibodies against specific cell surface targets.
- This method eliminates the need for antigen expression and purification, streamlining antibody generation.
- The technique is broadly applicable for developing targeted cancer immunotherapies.
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