title: Intravenous infusion of phage-displayed antibody library in human cancer patients: enrichment and cancer-specificity of tumor-homing phage-antibodies.
abstract: Phage display is a powerful method for target discovery and selection of ligands for cancer treatment and diagnosis. Our goal was to select tumor-binding antibodies in cancer patients. Eligibility criteria included absence of preexisting anti-phage-antibodies and a Stage IV cancer status. All patients were intravenously administered 1 × 10(11) TUs/kg of an scFv library 1 to 4 h before surgical resection of their tumors. No significant adverse events related to the phage library infusion were observed. Phage were successfully recovered from all tumors. Individual clones from each patient were assessed for binding to the tumor from which clones were recovered. Multiple tumor-binding phage-antibodies were identified. Soluble scFv antibodies were produced from the phage clones showing higher tumor binding. The tumor-homing phage-antibodies and derived soluble scFvs were found to bind varying numbers (0-5) of 8 tested normal human tissues (breast, cervix, colon, kidney, liver, spleen, skin, and uterus). The clones that showed high tumor-specificity were found to bind corresponding tumors from other patients also. Clone enrichment was observed based on tumor binding and DNA sequence data. Clone sequences of multiple variable regions showed significant matches to certain cancer-related antibodies. One of the clones (07-2,355) that was found to share a 12-amino-acid-long motif with a reported IL-17A antibody was further studied for competitive binding for possible antigen target identification. We conclude that these outcomes support the safety and utility of phage display library panning in cancer patients for ligand selection and target discovery for cancer treatment and diagnosis.
authors.affiliation: Department of Surgery, University of Vermont College of Medicine, Burlington, VT 05405, USA. Girja.Shukla@uvm.edu
authors.forename: Girja S
authors.initials: GS
authors.lastname: Shukla
authors.forename: David N
authors.initials: DN
authors.lastname: Krag
authors.forename: Elena N
authors.initials: EN
authors.lastname: Peletskaya
authors.forename: Stephanie C
authors.initials: SC
authors.lastname: Pero
authors.forename: Yu-Jing
authors.initials: YJ
authors.lastname: Sun
authors.forename: Chelsea L
authors.initials: CL
authors.lastname: Carman
authors.forename: Laurence E
authors.initials: LE
authors.lastname: McCahill
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Roland
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000912:Antibodies, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D005500:Follow-Up Studies; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007262:Infusions, Intravenous; D020381:Interleukin-17 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D009367:Neoplasm Staging; D009369:Neoplasms / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000912:Antibodies, Neoplasm; D007074:Immunoglobulin G; D020381:Interleukin-17; D019151:Peptide Library; D057127:Single-Chain Antibodies