Methods of assembling modified adenoviruses, libraries of adenoviral gene modules and compositions thereof are provided herein.

The present invention relates to a method for virus assay. More closely the invention relates a method for total quantification of adenovirus in a sample as well as total and functional (active) adenovirus in a sample. The method for determining adenovirus concentration in a sample comprises subjecting said sample to SPR (surface plasmon resonance) assay with immobilized FX (Factor X) and/or immobilized CAR (coxsackievirus and adenovirus receptor) on a sensor surface, wherein the adenovirus concentration is determined from sample binding to immobilized FX and/or immobilized CAR. CAR can be replaced by an ligand binding to adenovirus fiber, such as an anti-adenovirus fiber antibody. FX can be replaced by a ligand binding to adenovirus hexon, such as an anti-adenovirus hexon antibody. The method can be used for quality control in an adenovirus purification process, for example for gene therapy.

Recombinant adenovirus genomes that include an exogenous open reading frame (ORF) and a self-cleaving peptide coding sequence are described. Optimal placement of the exogenous genes for minimal impact on viral kinetics is further disclosed. Therapeutic applications of the recombinant adenoviruses are also described.

A method of purifying a composition comprising a group B adenovirus, for example comprising a purification step of: subjecting a composition comprising said group B adenovirus to diafiltration employing a diafiltration-buffer with a conductivity of at least 180 mScm.sup.−1, for example a conductivity of 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, or 290 mScm.sup.−1. Also provided is a composition obtained using the purification method disclosed herein.

A Process for the Production of Adenovirus The present disclosure relates to a continuous process for the manufacture of an adenovirus wherein the process comprises the steps: A) continuously culturing, in a vessel, mammalian cells infected with the adenovirus in the presence of media suitable for supporting the cells such that the virus replicates, wherein the cells are capable of supporting viral replication, and B) isolating from the media the virus produced from step a) wherein the isolation of virus is not subsequent to a cell lysis step, wherein viable cells for virus infection and production are maintained in the vessel at a level suitable for replicating the virus for the period of continuous manufacture, wherein the process comprises at least one media change or addition and at least one cell change or addition. The disclosure also extends to viruses populations obtained or obtainable from the method.

The present invention relates to a nucleotide sequence as shown in SEQ ID NO: 1 for encoding a Marburg virus envelope glycoprotein, and to a human replication-deficient recombinant adenovirus capable of expressing the nucleotide sequence and a preparation method therefor, as well as an application thereof in the preparation of a vaccine against Marburg virus disease. The vaccine uses an E1 and E3 deleted replication-deficient human type-5 adenovirus as a vector, and HEK293 cells integrating an adenovirus E1 gene as a packaging cell line, and a protective antigen gene carried is a codon-optimized Marburg virus Angola strain envelope glycoprotein gene. After codon optimization of the envelope glycoprotein gene, significant expression of envelope glycoprotein can be detected in transfected cells.

Embodiments herein relate to compositions of and methods for live viruses. In certain embodiments, a live, attenuated virus composition includes, but is not limited to, one or more live, attenuated viruses and compositions to reduce inactivation and/or degradation of the live, attenuated virus. In other embodiments, the live, attenuated virus composition may be a vaccine composition. In yet other compositions, a live, attenuated virus composition may include at least one carbohydrate, at least one protein and at least one high molecular weight surfactants for reducing inactivation and/or degradation of the live, attenuated virus.

The present invention relates to a method for preparing viral vector-based compositions wherein the viral vector-based particles present in the composition have a particle size distribution with a polydispersity index (PDI) of less than 0.5, the method comprising the steps: (a) providing replication-deficient viral vectors; (b) providing a solution comprising at least one sugar and at least three different excipients selected from hydrophilic and amphiphilic excipients, wherein the excipients are characterized by polar, aliphatic, aromatic, negatively charged, and/or positively charged functional groups, and wherein the solution is further characterized by an excipient-sugar ratio of at least 1:2 (w/w); and (c) mixing the replication deficient viral vectors of step (a) with the solution of step (b). The present invention further relates to a viral vector-based composition obtainable by the method of the invention as well as to the viral vector-based composition of the invention for use as a prime-boost vaccine.

A spiral tube countercurrent chromatography rotor for separating virus in a two part aqueous solvent is described.

Methods of purifying adenovirus that can be performed on a large scale. The methods purify adenovirus from an adenovirus-containing sample comprising or derived from a host cell population by clarifying the sample, wherein clarification comprises depth filtration followed by microfiltration; processing the clarified sample by anion exchange chromatography; and processing the anion exchange product by tangential flow filtration (TFF) to provide a TFF product.