Recombinant AAV Vectors for Enhanced Expression of Authentic IgG

dc.contributor.authorFuchs, Sebastian P.
dc.contributor.authorMartinez-Navio, José M.
dc.contributor.authorGao, Guangping
dc.contributor.authorDesrosiers, Ronald C.
dc.date.accessioned2016-08-16
dc.date.available2016-08-01
dc.date.created2016
dc.date.issued2016-08-16
dc.description.abstractAdeno-associated virus (AAV) has become a vector of choice for the treatment of a variety of genetic diseases that require safe and long-term delivery of a missing protein. Muscle-directed gene transfer for delivery of protective antibodies against AIDS viruses and other pathogens has been used experimentally in mice and monkeys. Here we examined a number of variations to AAV vector design for the ability to produce authentic immunoglobulin G (IgG) molecules. Expression of rhesus IgG from a single single-stranded AAV (ssAAV) vector (one vector approach) was compared to expression from two self-complementary AAV (scAAV) vectors, one for heavy chain and one for light chain (two vector approach). Both the one vector and the two vector approaches yielded considerable levels of expressed full-length IgG. A number of modifications to the ssAAV expression system were then examined for their ability to increase the efficiency of IgG expression. Inclusion of a furin cleavage sequence with a linker peptide just upstream of the 2A self-cleaving sequence from foot-and-mouth disease virus (F2A) increased IgG expression approximately 2 fold. Inclusion of these sequences also helped to ensure a proper sequence at the C-terminal end of the heavy chain. Inclusion of the post-transcriptional regulatory element from woodchuck hepatitis virus (WPRE) further increased IgG expression 1.5–2.0 fold. IgG1 versions of the two rhesus IgGs that were examined consistently expressed better than the IgG2 forms. In contrast to what has been reported for AAV2-mediated expression of other proteins, introduction of capsid mutations Y445F and Y731F did not increase ssAAV1-mediated expression of IgG as determined by transduction experiments in cell culture. Our findings provide a rational basis for AAV vector design for expression of authentic IgG.en
dc.format.extent19 Seiten
dc.identifier.citationPLoS ONE 11.6 (2016). <http://dx.doi.org/10.1371/journal.pone.0158009>
dc.identifier.opus-id7317
dc.identifier.urihttps://open.fau.de/handle/openfau/7317
dc.identifier.urnurn:nbn:de:bvb:29-opus4-73173
dc.language.isoen
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/de/deed.de
dc.subjectViral packaging
dc.subjectTransfection
dc.subjectEnzyme-linked immunoassays
dc.subjectPlasmid construction
dc.subjectCell cultures
dc.subjectPeptides
dc.subjectAntibodies
dc.subjectSequence motif analysis
dc.subject.ddcDDC Classification::6 Technik, Medizin, angewandte Wissenschaften :: 61 Medizin und Gesundheit :: 610 Medizin und Gesundheit
dc.titleRecombinant AAV Vectors for Enhanced Expression of Authentic IgGen
dc.typearticle
dcterms.publisherFriedrich-Alexander-Universität Erlangen-Nürnberg (FAU)
local.journal.issue6
local.journal.titlePLoS ONE
local.journal.volume11
local.sendToDnbfree*
local.subject.fakultaetMedizinische Fakultät
local.subject.gnd-
local.subject.sammlungUniversität Erlangen-Nürnberg / Open Access Artikel ohne Förderung / Open Access Artikel ohne Förderung 2016
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