title: Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody.
abstract: Therapy of melanoma using T-cells with genetically introduced T-cell receptors (TCRs) directed against a tumor-selective cancer testis antigen (CTA) NY-ESO1 demonstrated clear antitumor responses in patients without side effects. Here, we exploited the concept of TCR-mediated targeting through introduction of single-chain variable fragment (scFv) antibodies that mimic TCRs in binding major histocompatibility complex-restricted CTA. We produced scFv antibodies directed against Melanoma AntiGEn A1 (MAGE A1) presented by human leukocyte antigen A1 (HLA-A1), in short M1/A1, and coupled these TCR-like antibodies to liposomes to achieve specific melanoma targeting. Two anti-M1/A1 antibodies with different ligand-binding affinities were derived from a phage-display library and reformatted into scFvs with an added cysteine at their carboxyl termini. Protein production conditions, ie, bacterial strain, temperature, time, and compartments, were optimized, and following production, scFv proteins were purified by immobilized metal ion affinity chromatography. Batches of pure scFvs were validated for specific binding to M1/A1-positive B-cells by flow cytometry. Coupling of scFvs to liposomes was conducted by employing different conditions, and an optimized procedure was achieved. In vitro experiments with immunoliposomes demonstrated binding of M1/A1-positive B-cells as well as M1/A1-positive melanoma cells and internalization by these cells using flow cytometry and confocal microscopy. Notably, the scFv with nonenhanced affinity of M1/A1, but not the one with enhanced affinity, was exclusively bound to and internalized by melanoma tumor cells expressing M1/A1. Taken together, antigen-mediated targeting of tumor cells as well as promoting internalization of nanoparticles by these tumor cells is mediated by TCR-like scFv and can contribute to melanoma-specific targeting.
authors.affiliation: Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Mesha
authors.initials: M
authors.lastname: Saeed
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands.
authors.forename: Mandy
authors.initials: M
authors.lastname: van Brakel
authors.affiliation: Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Sara
authors.initials: S
authors.lastname: Zalba
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: Schooten
authors.affiliation: Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Joost A P
authors.initials: JA
authors.lastname: Rens
authors.affiliation: Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Gerben A
authors.initials: GA
authors.lastname: Koning
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands.
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
authors.affiliation: Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Timo L M
authors.initials: TL
authors.lastname: Ten Hagen
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D015788:HLA-A1 Antigen / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D008545:Melanoma / Q000188:drug therapy* / Q000276:immunology* / Q000378:metabolism; D058950:Melanoma-Specific Antigens / Q000276:immunology*; D058990:Molecular Targeted Therapy / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000008:administration & dosage* / Q000276:immunology* / Q000627:therapeutic use; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
chemical_list: D015788:HLA-A1 Antigen; D008081:Liposomes; C072011:MAGEA1 protein, human; D058950:Melanoma-Specific Antigens; D011948:Receptors, Antigen, T-Cell; D057127:Single-Chain Antibodies
keywords: HLA-A1 targeting; cancer; cancer testis antigens; immunotherapy; nanoparticles