title: Low cost delivery of proteins bioencapsulated in plant cells to human non-immune or immune modulatory cells.
abstract: Targeted oral delivery of GFP fused with a GM1 receptor binding protein (CTB) or human cell penetrating peptide (PTD) or dendritic cell peptide (DCpep) was investigated. Presence of GFP(+) intact plant cells between villi of ileum confirm their protection in the digestive system from acids/enzymes. Efficient delivery of GFP to gut-epithelial cells by PTD or CTB and to M cells by all these fusion tags confirm uptake of GFP in the small intestine. PTD fusion delivered GFP more efficiently to most tissues or organs than the other two tags. GFP was efficiently delivered to the liver by all fusion tags, likely through the gut-liver axis. In confocal imaging studies of human cell lines using purified GFP fused with different tags, GFP signal of DCpep-GFP was only detected within dendritic cells. PTD-GFP was only detected within kidney or pancreatic cells but not in immune modulatory cells (macrophages, dendritic, T, B, or mast cells). In contrast, CTB-GFP was detected in all tested cell types, confirming ubiquitous presence of GM1 receptors. Such low-cost oral delivery of protein drugs to sera, immune system or non-immune cells should dramatically lower their cost by elimination of prohibitively expensive fermentation, protein purification cold storage/transportation and increase patient compliance.
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Kwang-Chul
authors.initials: KC
authors.lastname: Kwon
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Brad E
authors.initials: BE
authors.lastname: Hoffman
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Aditya
authors.initials: A
authors.lastname: Kamesh
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Noah T
authors.initials: NT
authors.lastname: Jones
authors.affiliation: Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL, USA.
authors.forename: Roland W
authors.initials: RW
authors.lastname: Herzog
authors.affiliation: Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA. Electronic address: hdaniell@upenn.edu.
authors.forename: Henry
authors.initials: H
authors.lastname: Daniell
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D016503:Drug Delivery Systems / Q000191:economics; D005260:Female; D049452:Green Fluorescent Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D006801:Humans; D007107:Immune System / Q000166:cytology; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D059828:Plant Cells / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics
chemical_list: D001426:Bacterial Proteins; D057846:Cell-Penetrating Peptides; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C034993:ganglioside receptor; D049452:Green Fluorescent Proteins
keywords: Cell-penetrating peptides; Chloroplasts; Molecular farming; Protein drug delivery