The present invention pertains to methods and kits for enriching extracellular nucleic acids such as vesicular RNA from a sample comprising extracellular vesicles. Accordingly to the methods an acidic binding mixture is prepared comprising the sample and anion exchange particles and binding extracellular vesicles to the anion exchange particles. After separating the anion exchange particles comprising the bound extracellular vesicles from the remaining mixture, bound extracellular vesicles are lysing the in the presence of at least one detergent and released RNA is bound to the anion exchange particles. The anion exchange particles with the bound RNA from the lysate are then eluted.

Methods and systems for processing polynucleotides (e.g., DNA) are disclosed. A processing region includes one or more surfaces (e.g., particle surfaces) modified with ligands that retain polynucleotides under a first set of conditions (e.g., temperature and pH) and release the polynucleotides under a second set of conditions (e.g., higher temperature and/or more basic pH). The processing region can be used to, for example, concentrate polynucleotides of a sample and/or separate inhibitors of amplification reactions from the polynucleotides. Microfluidic devices with a processing region are disclosed.

The present disclosure pertains to methods of modifying an immune cell by delivering a modified messenger RNA (mRNA) encoding a chimeric antigen receptor (CAR) and modified immune cells comprising CARs.

A method is provided for enriching extracellular DNA from a biological sample comprising extracellular DNA and extracellular vesicles, wherein the method comprises: (a) preparing a binding mixture comprising—the biological sample, —a solid phase comprising anion exchange groups, —an acidic binding buffer comprising a buffering agent, and binding extracellular DNA to the solid phase comprising anion exchange groups; (b) separating the solid phase with the bound extracellular DNA from the remaining binding mixture, wherein the remaining binding mixture comprises extracellular vesicles. The method may furthermore comprise processing the remaining binding mixture to enrich one or more biological targets of interest therefrom, wherein processing may comprise (c) enriching as biological targets extracellular vesicles and/or extracellular RNA from the remaining binding mixture.

A porous membrane obtainable by a process comprising curing a composition comprising: (i) cross-linking agent(s) comprising at least one ligand group; (ii) inert solvent(s); (iii) polymerization initiator(s); and (vi) optionally monomer(s) other than component (i) which are reactive with component (i); wherein the composition satisfies the following equation: Z=wt(i)/(wt(i)+wt(iii)+wt(iv)) wherein: Z has a value of at least 0.6; wt(i) is the number of grammes of component (i) present in the composition; wt(iii) is the number of grammes of component (iii) present in the composition; and wt(iv) is the number of grammes of component (iv) present in the composition.

Separation media includes a membrane and a plurality of ligands immobilized on the membrane, the plurality of ligands comprising anion-exchange ligands, cation-exchange ligands, thiophilic ligands, hydrophilic ligands, hydrophobic interaction ligands, or a combination thereof. The separation media may be multimodal. The separation media may be configured for separation of target molecules comprising a nucleic acid, nucleotide, nucleoside, nucleobase, or an analogue or derivative thereof, from a reaction mixture. The separation media may be configured for use with organic solvents. A separation device includes the separation media. Materials including a nucleic acid, nucleotide, nucleoside, nucleobase, or an analogue or derivative thereof, may be purified at high speeds using the separation device.

A sample containing particles having high-molecular-weight (HMW) DNA is entrapped in a gel matrix, and the gel matrix is exposed to a lysis reagent configured to release the HMW DNA from the particles. The HMW DNA may be purified by subjecting the gel matrix to an electrophoretic field that removes the HMW DNA from the particles, lysis reagents, and/or other sample constituents, from the gel matrix such that the HMW DNA remains. The gel matrix may be subjected with DNA cleavase re-agents configured to cleave at specific DNA sequences within the HMW DNA to liberate defined segments of the DNA as fragments of reduced size. The gel matrix may also be subjected to an electrophoretic field, which moves and separates the DNA fragments from uncleaved DNA of the HMW DNA, which remains substantially immobile. The electrophoretically separated DNA fragments may be isolated from the gel matrix.

A test device is provided that can comprise: a housing accommodating a chromatography support, wherein the housing comprises: a supporting part that supports a container accommodating a liquid used for chromatography. A method is provided for performing chromatography using the test device.

Provided herein is a method to reduce the endotoxin contamination in plasmid preparations. In the described method, plasmid DNA and endotoxins are bound to an anionic exchange matrix and are brought into contact with a wash buffer, comprising a combination of branched secondary alcohol ethoxylates with varying ethylene oxide chain lengths, wherein the branched secondary alcohol ethoxylates with the shorter ethoxylate chain is present in the washing buffer in excess compared to the branched secondary alcohol ethoxylate with the longer ethoxylate chain. The resulting purified plasmid has minimal endotoxin contamination levels and is considered endotoxin-free. Furthermore, provided are wash buffers comprising a combination of branched secondary alcohol ethoxylates with varying ethylene oxide chain lengths, kits comprising such wash buffers and the use of such wash buffers for reducing the endotoxin contamination in plasmid preparations.

The present invention is directed to a method for modifying the retention time of RNA on a chromatographic column. The present invention also concerns a method for purifying RNA from a mixture of at least two RNA species. Furthermore, the present invention relates to a method for co-purifying at least two RNA species from a mixture of at least two RNA species. In particular, the present invention provides a method for harmonizing the numbers of A and/or U nucleotides in at least two RNA species. The present invention is also directed to RNA obtainable by said methods, a composition comprising said RNA or a vaccine comprising said RNA and methods for producing such RNA and compositions. Further, the invention concerns a kit, particularly a kit of parts, comprising the RNA, composition or vaccine. The invention is further directed to a method of treating or preventing a disorder or a disease, first and second medical uses of the RNA, composition and vaccine. Moreover, the present invention concerns a method for providing an adapted RNA sequence or an adapted RNA mixture.