Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

Mesha Saeed,1 Mandy van Brakel,2 Sara Zalba,1 Erik Schooten,2 Joost AP Rens,1 Gerben A Koning,1,&dagger; Reno Debets,2 Timo LM ten Hagen1 1Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands; 2Laboratory of Tumor...

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Autores principales:	Saeed M, van Brakel M, Zalba S, Schooten E, Rens JAP, Koning GA, Debets R, ten Hagen TLM
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2016
Materias:	Cancer Immunotherapy Nanoparticles Cancer Testis Antigens HLA-A1 targeting Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/818215a9953f41179a4d233311e1bcc6
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spelling	oai:doaj.org-article:818215a9953f41179a4d233311e1bcc62021-12-02T04:28:16ZTargeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody1178-2013https://doaj.org/article/818215a9953f41179a4d233311e1bcc62016-03-01T00:00:00Zhttps://www.dovepress.com/targeting-melanoma-with-immunoliposomes-coupled-to-anti-mage-a1-tcr-li-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Mesha Saeed,1 Mandy van Brakel,2 Sara Zalba,1 Erik Schooten,2 Joost AP Rens,1 Gerben A Koning,1,&dagger; Reno Debets,2 Timo LM ten Hagen1 1Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands; 2Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands &dagger;Dr Gerben A Koning passed away on December 29, 2015 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. Keywords: cancer, immunotherapy, nanoparticles, cancer testis antigens, HLA-A1 targetingSaeed Mvan Brakel MZalba SSchooten ERens JAPKoning GADebets Rten Hagen TLMDove Medical PressarticleCancerImmunotherapyNanoparticlesCancer Testis AntigensHLA-A1 targetingMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2016, Iss Issue 1, Pp 955-975 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cancer Immunotherapy Nanoparticles Cancer Testis Antigens HLA-A1 targeting Medicine (General) R5-920
spellingShingle	Cancer Immunotherapy Nanoparticles Cancer Testis Antigens HLA-A1 targeting Medicine (General) R5-920 Saeed M van Brakel M Zalba S Schooten E Rens JAP Koning GA Debets R ten Hagen TLM Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
description	Mesha Saeed,1 Mandy van Brakel,2 Sara Zalba,1 Erik Schooten,2 Joost AP Rens,1 Gerben A Koning,1,&dagger; Reno Debets,2 Timo LM ten Hagen1 1Laboratory of Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC, Rotterdam, the Netherlands; 2Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands &dagger;Dr Gerben A Koning passed away on December 29, 2015 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. Keywords: cancer, immunotherapy, nanoparticles, cancer testis antigens, HLA-A1 targeting
format	article
author	Saeed M van Brakel M Zalba S Schooten E Rens JAP Koning GA Debets R ten Hagen TLM
author_facet	Saeed M van Brakel M Zalba S Schooten E Rens JAP Koning GA Debets R ten Hagen TLM
author_sort	Saeed M
title	Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
title_short	Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
title_full	Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
title_fullStr	Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
title_full_unstemmed	Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody
title_sort	targeting melanoma with immunoliposomes coupled to anti-mage a1 tcr-like single-chain antibody
publisher	Dove Medical Press
publishDate	2016
url	https://doaj.org/article/818215a9953f41179a4d233311e1bcc6
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Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

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