Premium
Molecular construction of bionanoparticles: chimaeric SIV p17–HIV I p6 nanoparticles with minimal viral protein content
Author(s) -
Costa Maria J. L.,
Pedro Luísa,
De Matos António P. Alves,
AiresBarros Maria R.,
Belo José A.,
Goncalves João,
Ferreira Guilherme N. M.
Publication year - 2007
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20060208
Subject(s) - simian immunodeficiency virus , biology , virology , viral matrix protein , virus like particle , virus , vesicular stomatitis virus , viral protein , glycoprotein , hek 293 cells , microbiology and biotechnology , gene , biochemistry , recombinant dna
VLPs (virus‐like particles) are promising delivery vectors for molecular therapy, since they combine the major advantages of viral vectors with significantly fewer viral vector disadvantages. The present paper describes the molecular construction of chimaeric VLPs based on minimal SIV (simian immunodeficiency virus) and HIV1 components. A chimaeric protein was constructed by fusion of SIV matrix protein (p17) and HIV1 p6 protein, and we demonstrated that the chimaeric proteins assemble as 80 nm nanoparticles containing ∼7700 chimaeric protein units. Chimaeric VLPs are released from HEK‐293T cells (human embryonic kidney cells expressing the large T‐antigen of simian virus 40) and are fully encapsulated with lipid membrane. Chimaeric VLPs are produced at 3.7‐fold higher levels when compared with SIV p17 VLPs owing to duplication of a PTAP (Pro‐Thr‐Ala‐Pro) domain previously shown as essential for virus particle release. The chimaeric VLPs constructed in the present paper were efficiently pseudotyped with vesicular‐stomatitis‐virus glycoprotein, as shown by immunoprecipitation assays.