Provided herein are methods to characterize preparations of recombinant viral particles using analytical ultracentrifugation. Recombinant viral particles include recombinant adeno-associated viral particles, recombinant adenoviral particles, recombinant lentiviral particles and recombinant herpes simplex virus particles. Variant species of recombinant viral particles including empty capsids and recombinant viral particles with variant genomes (e.g., truncated genomes, aggregates, recombinants) can be identified and quantitated. The methods can be used to characterize preparations of recombinant viral particles regardless of the sequence of the recombinant viral genome or the serotype of the recombinant viral capsid.

Immunogenic compositions comprising viral vectors and surfactants are provided. Methods for administration and preparation of such compositions are also provided.

Vaccine compositions that may be administered to a subject via the buccal and/or sublingual mucosa are provided. Methods for administration and preparation of such vaccine compositions are also provided.

Viruses, and particularly genetically engineered, replication deficient viruses such as adenoviruses, poxviruses, MVA viruses, and baculoviruses which encode one or more antigens of interest, such as TB, malarial, and HIV antigens, are spray dried with a mannitol-cyclodextrin-trehalose-dextran (MCTD) to form a powder where the viability of the viruses are maintained at a suitable level for mass vaccinations after spray drying, and where the viability of the viruses are maintained at suitable level over a period of storage time, even in the presence of humidity.

Provided are compounds that enhance the efficacy of viruses by increasing spread of the virus in cells, increasing the titer of virus in cells, or increasing the antigen expression from a virus, gene or trans-gene expression from a virus, or virus protein expression in cells. Other uses, compositions and methods of using same are also provided.

Vaccine compositions that may be administered to a subject via the buccal and/or sublingual mucosa are provided. Methods for administration and preparation of such vaccine compositions are also provided.

Immunogenic compositions comprising viral vectors and surfactants are provided. Methods for administration and preparation of such compositions are also provided.

A process for assaying viral vector manufactured by large-scale viral vector manufacturing processes to assure the resulting vector has acceptable purity and potency. The process entails three different types of assays, each one of which is optionally useful on a stand-alone basis, and which together provide the first system able to assure the quality of viral vector produced by large-scale vector manufacturing processes,

A method for purifying a viral vector from a solution including the viral vector and host cell proteins (HCP) is provided. The method includes circulating the solution across an ultrafiltration/diafiltration membrane using tangential flow filtration (TFF) mode with continuous addition of diafiltration buffer. The method further includes filtering the solution across the ultrafiltration/diafiltration membrane to provide a permeate and a retentate. The retentate is collected such that a purified viral vector solution is obtained and retained in the retentate. A volume of the retentate is kept constant by the continuous addition of diafiltration buffer and the HCP is filtered out via the permeate.

The invention relates to methods for generating a recombinant adenovirus comprising a nucleotide sequence encoding a heterologous gene of interest for use as a vaccine comprising the steps of inserting the heterologous gene of interest into the adenovirus genome by recombining terminal protein complexed adenovirus genomic DNA (TPC-Ad gDNA) with a polynucleotide comprising a nucleotide sequence encoding the gene of interest and having 5′ and 3′ ends that are homologous to the insertion site sequence of the adenovirus genomic DNA in an in vitro recombination reaction, transfecting cells growing in individual vessels with a dilution of the in vitro recombination reaction mixture from (i) such that a number of such individual vessels contain a single cell that is infected by a recombinant adenovirus comprising the nucleotide sequence encoding the heterologous gene of interest, and identifying those individual vessels in which a single cell has been infected by the recombinant adenovirus comprising the nucleotide sequence encoding the heterologous gene of interest. Suitably said TPC-Ad gDNA comprises serotype-matched terminal protein and adenovirus genome, and said gene of interest codes for a single epitope, a string of epitopes, a segment of an antigen or a complete antigen protein. The invention also relates to recombinant adenoviruses and compositions made using these methods.