Multifunctional Transmembrane Protein Ligands for Cell-Specific Targeting of Plasma Membrane-Derived Vesicles.
Liposomes and nanoparticles that bind selectively to cell-surface receptors can target specific populations of cells. However, chemical conjugation of ligands to these particles is difficult to control, frequently limiting ligand uniformity and complexity. In contrast, the surfaces of living cells are decorated with highly uniform populations of sophisticated transmembrane proteins. Toward harnessing cellular capabilities, here it is demonstrated that plasma membrane vesicles (PMVs) derived from donor cells can display engineered transmembrane protein ligands that precisely target cells on the basis of receptor expression. These multifunctional targeting proteins incorporate (i) a protein ligand, (ii) an intrinsically disordered protein spacer to make the ligand sterically accessible, and (iii) a fluorescent protein domain that enables quantification of the ligand density on the PMV surface. PMVs that display targeting proteins with affinity for the epidermal growth factor receptor (EGFR) bind at increasing concentrations to breast cancer cells that express increasing levels of EGFR. Further, as an example of the generality of this approach, PMVs expressing a single-domain antibody against green fluorescence protein (eGFP) bind to cells expressing eGFP-tagged receptors with a selectivity of ≈50:1. The results demonstrate the versatility of PMVs as cell targeting systems, suggesting diverse applications from drug delivery to tissue engineering.
Department of Biomedical Engineering, Institute for Cellular and Molecular Biology, The University of Texas at Austin, TX, 78712, USA.
Chi
C
Zhao
Department of Biomedical Engineering, Institute for Cellular and Molecular Biology, The University of Texas at Austin, TX, 78712, USA.
David J
DJ
Busch
Department of Biomedical Engineering, Institute for Cellular and Molecular Biology, The University of Texas at Austin, TX, 78712, USA.
Connor P
CP
Vershel
Department of Biomedical Engineering, Institute for Cellular and Molecular Biology, The University of Texas at Austin, TX, 78712, USA.
Jeanne C
JC
Stachowiak
D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism*; D003412:Cricetulus; D016503:Drug Delivery Systems; D066246:ErbB Receptors / Q000737:chemistry; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D023822:Tissue Engineering
D008081:Liposomes; D066246:ErbB Receptors
biomaterials; cell targeting; extracellular vesicles; protein engineering