A method for separating full Adeno-associated virus (AAV) capsids from empty AAV capsids in a buffered mixture comprising full AAV capsids, empty AAV capsids, comprising the steps of contacting the buffered mixture with a first substrate bearing a metal affinity ligand attached to the first substrate, said metal affinity ligand having the ability to complex metal ions via three or more nitrogen atoms, separating empty AAV capsids from full AAV capsids by eluting with a pH gradient, a salt gradient, a metal ion gradient or a combination thereof in the presence of multivalent cations bound to the metal affinity ligand to obtain a purified full AAV capsid fraction.

For removing contaminating DNA in the mixture or purified AAV capsid fraction, the method of the invention can be combined with contacting of the buffered mixture or the purified full AAV capsid fraction with a second substrate bearing a metal affinity ligand attached to the second substrate in the presence of multivalent cations bound to the metal affinity ligand, said metal affinity ligand comprises two or more negatively charged carboxylic acid residues.

Substantially pure saponin extracts and orthogonal chromatographic methods for purification of saponin extracts are disclosed. The purified saponin extracts may include QS-21 and can have a purity of greater than 97%. The orthogonal chromatographic method uses reversed-phase (RP) chromatography followed by hydrophilic interaction liquid chromatography (HILIC) to generate substantially pure saponin extracts.

The present disclosure relates to multistep chromatographic methods for preparing extracellular vesicles (EVs). The methods were demonstrated to be effective in preparing high quality EVs in a large scale. The methods enable preparation of EVs for therapeutic and diagnostic applications, and isolation and/or sub-fractionation of EVs with desired properties for specific use.

The present disclosure is directed to liquid chromatography columns and methods utilizing a stationary phase sorbent having two or more gradient factors. Each of the two or more gradient factors varies in a progressive manner along a length of the stationary phase sorbent in a direction from an inlet to an outlet of the column (i.e., along a length of the column). As a result of including these new continuous stationary phase gradients, new selectivities allowing for separation and analysis of complex samples including large biomolecules is achievable.

A system for preparing solutions for chromatography application is disclosed. The system comprises a T-joint for preparing a buffer solution by mixing at least one first solution and a second solution. The T-joint receives the second solution from a solution supply unit connected to the T-joint. Further one or more low pressure pumps supply the one or more first solutions into the T-joint. The high pressure pump collects the buffer solution and delivers it to a chromatography apparatus.

Disclosed is a gradient proportioning valve for liquid chromatography that includes a plurality of inlet ports configured to receive a plurality of fluids, a manifold connected to each of the plurality of inlet ports configured to mix the plurality of fluids in a controlled manner to provide a fluid composition, the manifold including a plurality of fluid conduits internal to the manifold, each of the plurality of fluid conduits receiving fluid through a respective one of the plurality of inlet ports, each of the plurality of fluid conduits operatively communicable to a respective actuation mechanism configured to open and close each of the plurality of fluid conduits in a controlled manner, a common outlet port configured to receive the fluid composition, and a passive fluidic dampening system configured to dampen unwanted fluidic pressure pulses in the manifold where at least one of the plurality of fluid conduits is compliant.

Disclosed is a gradient proportioning valve for use in liquid chromatography that includes a plurality of inlet ports configured to receive a plurality of fluids, a manifold connected to each of the plurality of inlet ports configured to mix the plurality of fluids in a controlled manner to provide a fluid composition, the manifold including a plurality of conduits internal to the manifold, each of the plurality of conduits receiving fluid through a respective one of the plurality of inlet ports, each of the plurality of conduits operatively communicable to a respective actuation mechanism configured to open and close each of the plurality of conduits in a controlled manner, a common outlet port configured to receive the fluid composition, and an active fluidic dampening system configured to dampen unwanted fluidic pressure pulses in the manifold. Liquid chromatography systems and methods are further disclosed.

A method is disclosed for manufacturing a purified pDNA preparation from a sample comprising pDNA and a contaminant, the method comprising the steps of: Contacting the sample with a hydrophobic interaction chromatography (HIC) material in a solution comprising a kosmotropic salt in a concentration which forces the pDNA and contaminant to adsorb on the HIC material, Diluting the concentration of the kosmotropic salt in presence of a neutral salt subsequent to adsorbing of the pDNA on the HIC material, thereby Desorbing the pDNA from the HIC material, whereas the contaminant stays adsorbed by the continued presence of the neutral salt, and Obtaining the pDNA preparation.

Furthermore, a method is disclosed for preparing a sample to be subjected to the method of the invention or other purification methods, in particular anion exchange chromatography by exposing the sample to a neutral salt in the presence of a HIC material.

Examples include microfluidic devices. Example microfluidic devices include a first microfluidic channel, a second microfluidic channel, and a third microfluidic channel fluidly coupled to the first microfluidic channel and the second microfluidic channel via a fluid junction. A fluid actuator is disposed in the third microfluidic channel to actuate to thereby pump a first fluid and a second fluid into the third microfluidic channel.

The present disclosure relates to the determination of analytes in a sample using chromatography. The present disclosure provides methods of separating an analyte from a sample. A mobile phase is flowed through a chromatography column. The mobile phase includes about 0.005% (v/v) to about 0.20% (v/v) difluoroacetic acid and less than about 100 ppb of any individual metal impurity. A sample including the analyte is injected into the mobile phase. The analyte is separated from the sample.