title: Anti-CD20 single chain variable antibody fragment-apolipoprotein A-I chimera containing nanodisks promote targeted bioactive agent delivery to CD20-positive lymphomas.
abstract: A fusion protein comprising an α-CD20 single chain variable fragment (scFv) antibody, a spacer peptide, and human apolipoprotein (apo) A-I was constructed and expressed in Escherichia coli. The lipid interaction properties intrinsic to apoA-I as well as the antigen recognition properties of the scFv were retained by the chimera. scFv•apoA-I was formulated into nanoscale reconstituted high-density lipoprotein particles (termed nanodisks; ND) and incubated with cultured cells. α-CD20 scFv•apoA-I ND bound to CD20-positive non-Hodgkins lymphoma (NHL) cells (Ramos and Granta) but not to CD20-negative T lymphocytes (i.e., Jurkat). Binding to NHL cells was partially inhibited by pre-incubation with rituximab, a monoclonal antibody directed against CD20. Confocal fluorescence microscopy analysis of Granta cells following incubation with α-CD20 scFv•apoA-I ND formulated with the intrinsically fluorescent hydrophobic polyphenol, curcumin, revealed α-CD20 scFv•apoA-I localizes to the cell surface, while curcumin off-loads and gains entry to the cell. Compared to control incubations, viability of cultured NHL cells was decreased upon incubation with α-CD20 scFv•apoA-I ND harboring curcumin. Thus, formulation of curcumin ND with α-CD20 scFv•apoA-I as the scaffold component confers cell targeting and enhanced bioactive agent delivery, providing a strategy to minimize toxicity associated with chemotherapeutic agents.
authors.affiliation: a Lypro Biosciences Inc., 1236 Hawthorne St. Alameda, CA 94501, USA.
authors.forename: Natasha M
authors.initials: NM
authors.lastname: Crosby
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Mistuni
authors.initials: M
authors.lastname: Ghosh
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Betty
authors.initials: B
authors.lastname: Su
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Beckstead
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Ayako
authors.initials: A
authors.lastname: Kamei
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Jens B
authors.initials: JB
authors.lastname: Simonsen
authors.affiliation: a Lypro Biosciences Inc., 1236 Hawthorne St. Alameda, CA 94501, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Luo
authors.affiliation: c Division of Hematology/Oncology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
authors.forename: Leo I
authors.initials: LI
authors.lastname: Gordon
authors.affiliation: a Lypro Biosciences Inc., 1236 Hawthorne St. Alameda, CA 94501, USA.|b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Trudy M
authors.initials: TM
authors.lastname: Forte
authors.affiliation: b Children's Hospital Oakland Research Institute, 5700 Martin Luther King Jr. Way, Oakland, CA 94609, USA.
authors.forename: Robert O
authors.initials: RO
authors.lastname: Ryan
mesh_terms: D018951:Antigens, CD20 / Q000276:immunology*; D016632:Apolipoprotein A-I / Q000276:immunology*; D045744:Cell Line, Tumor; D006801:Humans; D008223:Lymphoma / Q000276:immunology / Q000628:therapy*; D018613:Microscopy, Confocal; D049329:Nanostructures; D011993:Recombinant Fusion Proteins / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology*
chemical_list: D018951:Antigens, CD20; D016632:Apolipoprotein A-I; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies
keywords: CD-20; CD20; apolipoprotein A-I; apolipoprotéine A-I; confocal fluorescence microscopy; curcumin; curcumine; fragment de la chaîne variable simple d’un anticorps; lymphome non-Hodgkinien; microscopie confocale à fluorescence; nanodisk; nanodisque; non-Hodgkin’s lymphoma; single chain variable antibody fragment