دورية أكاديمية
High-content analysis of antibody phage-display library selection outputs identifies tumor selective macropinocytosis-dependent rapidly internalizing antibodies.
| العنوان: | High-content analysis of antibody phage-display library selection outputs identifies tumor selective macropinocytosis-dependent rapidly internalizing antibodies. |
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| المؤلفون: | Ha KD; From the ‡Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California 94110-1305., Bidlingmaier SM; From the ‡Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California 94110-1305., Zhang Y; From the ‡Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California 94110-1305., Su Y; From the ‡Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California 94110-1305., Liu B; From the ‡Department of Anesthesia, UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California 94110-1305 liub@anesthesia.ucsf.edu. |
| المصدر: | Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2014 Dec; Vol. 13 (12), pp. 3320-31. Date of Electronic Publication: 2014 Aug 22. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 101125647 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1535-9484 (Electronic) Linking ISSN: 15359476 NLM ISO Abbreviation: Mol Cell Proteomics Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Original Publication: Bethesda, MD : American Society for Biochemistry and Molecular Biology, [2002- |
| مواضيع طبية MeSH: | Peptide Library*, Antibodies, Neoplasm/*pharmacology , Antigens, Neoplasm/*immunology , Antineoplastic Agents/*pharmacology , Immunoglobulin G/*pharmacology , Receptor, EphA2/*immunology, Antibodies, Neoplasm/immunology ; Antibodies, Neoplasm/metabolism ; Antibody Affinity ; Antibody Specificity ; Antigen-Antibody Complex/genetics ; Antigen-Antibody Complex/immunology ; Antigen-Antibody Complex/metabolism ; Antigens, Neoplasm/genetics ; Antigens, Neoplasm/metabolism ; Antineoplastic Agents/immunology ; Antineoplastic Agents/metabolism ; Biomarkers/metabolism ; Cell Line ; Cell Line, Tumor ; Fibroblasts/cytology ; Fibroblasts/drug effects ; Fibroblasts/metabolism ; Gene Expression ; HEK293 Cells ; High-Throughput Screening Assays ; Humans ; Immunoglobulin G/immunology ; Immunoglobulin G/metabolism ; Immunotoxins/chemistry ; Immunotoxins/immunology ; Laser Capture Microdissection ; Molecular Targeted Therapy ; Pinocytosis ; Receptor, EphA2/genetics ; Receptor, EphA2/metabolism ; Ribosome Inactivating Proteins, Type 1/chemistry ; Ribosome Inactivating Proteins, Type 1/immunology ; Saporins |
| مستخلص: | Many forms of antibody-based targeted therapeutics, including antibody drug conjugates, utilize the internalizing function of the targeting antibody to gain intracellular entry into tumor cells. Ideal antibodies for developing such therapeutics should be capable of both tumor-selective binding and efficient endocytosis. The macropinocytosis pathway is capable of both rapid and bulk endocytosis, and recent studies have demonstrated that it is selectively up-regulated by cancer cells. We hypothesize that receptor-dependent macropinocytosis can be achieved using tumor-targeting antibodies that internalize via the macropinocytosis pathway, improving potency and selectivity of the antibody-based targeted therapeutic. Although phage antibody display libraries have been utilized to find antibodies that bind and internalize to target cells, no methods have been described to screen for antibodies that internalize specifically via macropinocytosis. We hereby describe a novel screening strategy to identify phage antibodies that bind and rapidly enter tumor cells via macropinocytosis. We utilized an automated microscopic imaging-based, High Content Analysis platform to identify novel internalizing phage antibodies that colocalize with macropinocytic markers from antibody libraries that we have generated previously by laser capture microdissection-based selection, which are enriched for internalizing antibodies binding to tumor cells in situ residing in their tissue microenvironment (Ruan, W., Sassoon, A., An, F., Simko, J. P., and Liu, B. (2006) Identification of clinically significant tumor antigens by selecting phage antibody library on tumor cells in situ using laser capture microdissection. Mol. Cell. Proteomics. 5, 2364-2373). Full-length human IgG molecules derived from macropinocytosing phage antibodies retained the ability to internalize via macropinocytosis, validating our screening strategy. The target antigen for a cross-species binding antibody with a highly active macropinocytosis activity was identified as ephrin type-A receptor 2. Antibody-toxin conjugates created using this macropinocytosing IgG were capable of potent and receptor-dependent killing of a panel of EphA2-positive tumor cell lines in vitro. These studies identify novel methods to screen for and validate antibodies capable of receptor-dependent macropinocytosis, allowing further exploration of this highly efficient and tumor-selective internalization pathway for targeted therapy development. (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.) |
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| معلومات مُعتمدة: | R01 CA118919 United States CA NCI NIH HHS; R01 CA129491 United States CA NCI NIH HHS; R01 CA171315 United States CA NCI NIH HHS |
| المشرفين على المادة: | 0 (Antibodies, Neoplasm) 0 (Antigen-Antibody Complex) 0 (Antigens, Neoplasm) 0 (Antineoplastic Agents) 0 (Biomarkers) 0 (Immunoglobulin G) 0 (Immunotoxins) 0 (Peptide Library) 0 (Ribosome Inactivating Proteins, Type 1) EC 2.7.10.1 (Receptor, EphA2) EC 3.2.2.22 (Saporins) |
| تواريخ الأحداث: | Date Created: 20140824 Date Completed: 20150710 Latest Revision: 20220410 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC4256486 |
| DOI: | 10.1074/mcp.M114.039768 |
| PMID: | 25149096 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1535-9484 |
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| DOI: | 10.1074/mcp.M114.039768 |