Plant viral elongated nanoparticles modified for log-increases of foreign peptide immunogenicity and specific antibody detection.
Elongated and flexuous recombinant nanoparticles were derived from Turnip mosaic virus to be used as bioscaffolds for increased peptide immunogenicity and peptide-specific antibody sensing. For this purpose, a 20-amino acid peptide derived from human vascular endothelial growth factor receptor 3 (VEGFR-3) was fused to the N-terminal region of Turnip mosaic virus coat protein (CP) by genetic insertion. The insertion was between codons corresponding to the first and second amino acids of the CP in two versions of a previously reported virus-derived vector. Systemic infections of two genetic constructs were achieved in two different plant hosts. The construct proved stable upon successive passages and generated virus nanoparticles identifiable under the electron microscope. The chimeric structures held the VEGFR-3 peptide. Purified VER3 nanoparticles were used to immunize mice, whose sera showed log increases of antibodies against the VEGFR-3 peptide when compared with mice immunized with peptide alone, thus providing the first quantitative data on the potential of elongated flexuous viruses for peptide immunogenicity increases. Purified VER3 nanoparticles also showed log increases in their ability to detect VER3 antibodies in sera, when used as reagents in ELISA assays, an application also used here for the first time.
Centro de Biotecnología y Genómica de Plantas (CBGP, UPM-INIA), Campus de Montegancedo, Autovía M40, Km 38, 28223 Pozuelo de Alarcón, Madrid, Spain.
Flora
F
Sánchez
María
M
Sáez
Pablo
P
Lunello
Fernando
F
Ponz
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D029688:Brassica napus / Q000821:virology; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017806:Tymovirus / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics* / Q000276:immunology; D014771:Virion / Q000235:genetics* / Q000276:immunology
D000906:Antibodies; D010455:Peptides; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2
Antibody; Detection; Peptide presentation; VEGFR-3; Viral nanoparticles