Anti-CD20 single chain variable antibody fragment-apolipoprotein A-I chimera containing nanodisks promote targeted bioactive agent delivery to CD20-positive lymphomas

Natasha M. Crosby, Mistuni Ghosh, Betty Su, Jennifer A. Beckstead, Ayako Kamei, Jens B. Simonsen, Bing Luo, Leo I Gordon, Trudy M. Forte, Robert O. Ryan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)343-350
Number of pages8
JournalBiochemistry and Cell Biology
Volume93
Issue number4
DOIs
StatePublished - Apr 20 2015

Keywords

  • Apolipoprotein A-I
  • CD20
  • Confocal fluorescence microscopy
  • Curcumin
  • Nanodisk
  • Non-Hodgkin's lymphoma
  • Single chain variable antibody fragment

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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