Abstract: Provided herein are methods of measuring the qualitative and/or quantity attributes of gene therapy vector preparations. In certain embodiments, the gene therapy vector preparations are AAV preparations (e.g., AAV8). In certain embodiments the methods comprise determining the potency or dose of the AAV preparation using ELISA or ELISA in combination with CryoTEM.

Abstract: A scalable process and device for producing a biomolecule, in particular pharmaceutical grade plasmid DNA. The process includes the steps of alkaline lysis and a neutralization. For separating the lysate and the precipitate, the mixture is allowed to gently flow downward through a clarification reactor that is partially filled, in its lower part, with retention material like glass beads, whereby the precipitate is retained on top of and within the retention. In a preferred embodiment of the lysis step, cell suspension and alkaline lysis solution flow through a lysis reactor that is filled with particulate material like glass beads. The process can be run continuously and fully automated.

Abstract: A scalable process and device for producing a biomolecule, in particular pharmaceutical grade plasmid DNA. The process includes the steps of alkaline lysis and a neutralization. For separating the lysate and the precipitate, the mixture is allowed to gently flow downward through a clarification reactor that is partially filled, in its lower part, with retention material like glass beads, whereby the precipitate is retained on top of and within the retention. In a preferred embodiment of the lysis step, cell suspension and alkaline lysis solution flow through a lysis reactor that is filled with particulate material like glass beads. The process can be run continuously and fully automated.

Abstract: A method for producing plasmid DNA on a manufacturing scale uses Escherichia coli K-12 strain JM108. The process results in high yield and homogeneity of plasmid DNA.

Abstract: In a method for reconstituting a recombinant protein from a denatured state to its active form, a feed solution containing the recombinant protein in its denatured and/or in biologically inactive intermediate forms is subjected to a chromatographic separation process, in which the protein is reconstituted under conditions that promote refolding of the protein and the intermediate forms are separated from the refolded protein. The denatured form and/or the inactive intermediate forms of the protein are separated from the refolded protein in a continuous or quasi-continuous manner and optionally recycled to the feed solution.

Abstract: A method for producing plasmid DNA on a manufacturing scale uses Escherichia coli K-12 strain JM108. The process results in high yield and homogeneity of plasmid DNA.

Abstract: The invention relates to a process for preparing pyroGlu-MCP-1 from recombinantly produced Gln-MCP-1, wherein Gln-MCP-1 is incubated at a temperature in the range from 30° C. and 80° C. in a buffer solution with a salt concentration in the range from 10 mM to 160 mM and a pH in the range from 2 to 7.5, until at least 90% of the MCP-1 is present in the form of the pyroGlu-MCP-1.

Abstract: A scalable process and device for producing a biomolecule, in particular pharmaceutical grade plasmid DNA. The process includes the steps of alkaline lysis and a neutralization. For separating the lysate and the precipitate, the mixture is allowed to gently flow downward through a clarification reactor that is partially filled, in its lower part, with retention material like glass beads, whereby the precipitate is retained on top of and within the retention. In a preferred embodiment of the lysis step, cell suspension and alkaline lysis solution flow through a lysis reactor that is filled with particulate material like glass beads. The process can be run continuosly and fully automated.

Abstract: The invention relates to a process for isolating and purifying a polynucleotide on a manufacturing scale which uses a chromatographic separation process comprising a combination of two different chromatographic steps selected from hydrophobic interaction chromatography, polar interaction chromatography and anion exchange chromatography. In at least one of the two steps the chromatographic support is a porous monolithic bed. The invention also relates to a chromatographic device and its use for isolating and purifying a polynucleotide of interest, in particular plasmid DNA, on manufacturing scale.

Abstract: In a method for reconstituting a recombinant protein from a denatured state to its active form, a feed solution containing the recombinant protein in its denatured and/or in biologically inactive intermediate forms is subjected to a chromatographic separation process, in which the protein is reconstituted under conditions that promote refolding of the protein and the intermediate forms are separated from the refolded protein. The denatured form and/or the inactive intermediate forms of the protein are separated from the refolded protein in a continuous or quasi-continuous manner and optionally recycled to the feed solution.