title: Cry protein crystals: a novel platform for protein delivery.
abstract: Protein delivery platforms are important tools in the development of novel protein therapeutics and biotechnologies. We have developed a new class of protein delivery agent based on sub-micrometer-sized Cry3Aa protein crystals that naturally form within the bacterium Bacillus thuringiensis. We demonstrate that fusion of the cry3Aa gene to that of various reporter proteins allows for the facile production of Cry3Aa fusion protein crystals for use in subsequent applications. These Cry3Aa fusion protein crystals are efficiently taken up and retained by macrophages and other cell lines in vitro, and can be delivered to mice in vivo via multiple modes of administration. Oral delivery of Cry3Aa fusion protein crystals to C57BL/6 mice leads to their uptake by MHC class II cells, including macrophages in the Peyer's patches, supporting the notion that the Cry3Aa framework can be used to stabilize cargo protein against degradation for delivery to gastrointestinal lymphoid tissues.
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Manoj S
authors.initials: MS
authors.lastname: Nair
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Marianne M
authors.initials: MM
authors.lastname: Lee
authors.affiliation: Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Astrid
authors.initials: A
authors.lastname: Bonnegarde-Bernard
authors.affiliation: Department of Molecular and Cellular Biology, The Ohio State University, Columbus, OH, United States of America; Tumor Microenvironment Program, The Comprehensive Cancer Center, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Wallace
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Donald H
authors.initials: DH
authors.lastname: Dean
authors.affiliation: Department of Molecular and Cellular Biology, The Ohio State University, Columbus, OH, United States of America; Tumor Microenvironment Program, The Comprehensive Cancer Center, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Ostrowski
authors.affiliation: Department of Neuroscience, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Burry
authors.affiliation: Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Prosper N
authors.initials: PN
authors.lastname: Boyaka
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, United States of America.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Chan
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D003460:Crystallization; D004731:Endotoxins / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006460:Hemolysin Proteins / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D008264:Macrophages / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D010581:Peyer's Patches / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D000067996:RAW 264.7 Cells; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D049452:Green Fluorescent Proteins; D008156:Luciferases