Methods of maintaining narrow residence time distributions in continuous flow systems, particularly applicable to virus inactivation such as during a protein purification process. Fluid sample is introduced into an axial flow channel and caused to flow therein in discrete packets or zones to minimize residence time distribution and axial dispersion. Embodiments described herein obviate or minimize the need for using large tanks or reservoirs for performing virus inactivation during a protein purification process; reduce the overall time required for virus inactivation, and/or reduce the overall physical space required to perform the virus inactivation operation during a protein purification process, which in turn reduces the overall footprint for the purification process.

The present disclosure relates to an enhanced multi-dimensional chromatography system and method using selectable At-Column Dilution to improve compatibility of the interface and transfer between the multiple dimensions. The use of At-Column Dilution (ACD) with multi-dimensional chromatography can provide greater retention of the diverted components on subsequent stationary phases, and increase the sensitivity and peak shape of the component(s) separated on subsequent dimensions.

Methods of maintaining narrow residence time distributions in continuous flow systems, particularly applicable to virus inactivation such as during a protein purification process. Fluid sample is introduced into an axial flow channel and caused to flow therein in discrete packets or zones to minimize residence time distribution and axial dispersion. Embodiments described herein obviate or minimize the need for using large tanks or reservoirs for performing virus inactivation during a protein purification process; reduce the overall time required for virus inactivation, and/or reduce the overall physical space required to perform the virus inactivation operation during a protein purification process, which in turn reduces the overall footprint for the purification process.

Provided herein are purification, production and manufacturing methods for recombinant viral vector particles such as recombinant adeno-associated viral (rAAV) vector particles substantially free of empty viral particles; a population of recombinant adeno-associated vims (rAAV) particles purified using the method described herein, and a pharmaceutical composition comprising the purified rAAV.

Provided is a system comprising a device that performs one or more reactions to liquid or supercritical fluid chromatograph effluents and produces molecules that are subsequently detected by a suitable detector. This allows for one to practice a method for the detection and quantification of organic molecules from a liquid chromatograph for the purpose of increasing detection limits and allowing for the universal detection of organic molecules. The linear dynamic range and molecular response are greater than those previously available.

A method for obtaining a liquid from a porous solid phase is described. The method comprises forming a liquid seal at a first end of a porous solid phase to which a liquid is bound, wherein liquid of the liquid seal is immiscible with the liquid bound to the solid phase, and applying a pressure differential across the porous solid phase to cause the immiscible liquid to move through the porous solid phase towards a second end of the porous solid phase, thereby displacing the liquid bound to the porous solid phase towards the second end and releasing this liquid from the second end. Recovery of liquid from the solid phase using such methods is increased compared with corresponding methods in which no liquid seal is formed. In preferred embodiments, the liquid used to form the liquid seal is a mineral oil. The methods have particular application in nucleic acid extractions which utilise capture of nucleic acid to a solid phase. Kits and apparatus for performing the methods are also described.

The present disclosure relates to an enhanced multi-dimensional chromatography system and method using selectable At-Column Dilution to improve compatibility of the interface and transfer between the multiple dimensions. The use of At-Column Dilution (ACD) with multi-dimensional chromatography can provide greater retention of the diverted components on subsequent stationary phases, and increase the sensitivity and peak shape of the component(s) separated on subsequent dimensions.

Provided is a system comprising a device that performs one or more reactions to liquid or supercritical fluid chromatograph effluents and produces molecules that are subsequently detected by a suitable detector. This allows for one to practice a method for the detection and quantification of organic molecules from a liquid chromatograph for the purpose of increasing detection limits and allowing for the universal detection of organic molecules. The linear dynamic range and molecular response are greater than those previously available.

A chromatographic purification method for the isolation of a product from a feed mixture using two columns, wherein in one step b) the first upstream column is loaded with feed, followed by at least one interconnected step c), wherein in the interconnected step c) the first column is fed with eluent with a gradient, wherein the stream exiting the first column is adjusted inline with inline adjustment eluent before entering the second column during the period of gradient elution, and wherein the inline adjustment eluent is as the eluent fed at the first column inlet but controlled to have a higher or lower modifier concentration than the eluent exiting at the first column, and wherein the modifier difference of the inline adjustment eluent is chosen such that the adherence of the product to the stationary phase of the second column is higher than without that difference.

Methods of mass spectrometry comprise providing a target list of mass to charge ratios of fragments of interest for the sample to be analysed that is ionized to form sample ions. A plurality of MS2 analyses across the m/z range of interest is performed, each MS2 analysis comprising mass selecting the sample ions using an isolation window having a first m/z width and fragmenting the sample ions within the isolation window. A centre m/z of the isolation window is updated such that the plurality of MS2 analyses cover the m/z range of interest. Each MS2 analysis also comprises mass analysing the fragment ions and determining a m/z associated with each spectral peak, and comparing the m/z of the spectral peaks to the m/z of the fragments of the target list. Upon detecting a match, a triggered MS2 analysis is performed with a higher sensitivity.