There is provided a system for performing a chromatographic separation of an analyte, methods of using the system to separate at least one component of an analyte and an eluent generator of use in the system. An exemplary system comprises: (a) an eluent generator comprising: (i) a housing configured to be pressurizable by gas, comprising an annular void defined by the housing, and a gas inlet for the gas and a gas outlet for the gas in fluid communication with the annular void; (ii) a membrane permeable to the gas defining an eluent flow channel disposed within the annular void, the eluent flow channel having an eluent precursor fluid inlet and an eluent outlet; (iii) a source of gas in fluidic communication with the gas inlet; (iv) a source of the eluent precursor fluid; and (b) a chromatography column disposed downstream of and in fluidic communication with the eluent outlet.

The present invention provides methods for isolating an active polypeptide or immunoconjugate by purification of a solution containing both the active polypeptide or immunoconjugate and an acidic variant thereof, such as a deamidated variant, using anion exchange chromatography. The present invention also provides compositions, formulations, and unit dosage forms comprising the purified polypeptide or immunoconjugate.

Provided herein are methods for the purification of recombinant Adeno-Associated Virus (rAAV) particles using anion exchange chromatography.

Disclosed are methods for identification of one or more non-native disulfide bonds in a biomolecule (e.g, an antibody). In an example, a method includes performing a digestion of the biomolecule under non-reducing conditions to provide a sample comprising a plurality of biomolecule fragments, contacting the sample to a separation column, applying a first mobile phase gradient comprising trifluoroacetic acid (TFA) and a small molecule additive to the separation column, applying a second mobile phase gradient comprising TFA in acetonitrile (ACN) and a small molecule additive to the separation column, performing a partial reduction procedure on the eluted sample, applying the partially reduced eluted sample components to a mass spectrometer, and performing a mass spectrometric analysis on the partially reduced eluted sample components to identify the one or more non-native disulfide bonds in the biomolecule.

The invention relates to a chromatography method of producing a target elution volume comprising a first and a second target protein. The method includes providing a cation exchange chromatography column; applying a protein solution on the column, the protein solution comprising the first target protein, a second target protein and optionally one or more further proteins; inputting an optimization criterion; computing chromatography simulations for computing an elution buffer salt concentration adapted to provide a target elution volume matching the optimization criterion best; computing the pooling borders of the target elution volume as a function of at least the computed salt concentration and the input optimization criterion; applying an elution buffer having the computed salt concentration on the chromatography column; performing the elution; and collecting the computed target elution volume.

A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.

A method of separating a liquid sample containing viral vectors includes flowing the liquid sample into an anion exchange column. A first viral vector contains a targeted genetic material and a second viral vector contains essentially no genetic material or a non-targeted genetic material, wherein the targeted genetic material is different than the non-targeted genetic material. A mobile phase can be flowed into the anion exchange column, wherein the mobile phase includes a buffer solution A and a buffer solution B. The buffer solutions A and B both include volatile buffer salts. The first viral vector and the second viral vector can be separated so that the first viral vector and the second viral vector elute off the anion exchange column at different times and then be detected.

The present disclosure provides methods for the separation and quantification of empty and full viral capsids (e.g., AAV capsids) within a viral preparation, such as a viral pharmaceutical composition or drug product.

A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.

A lamp-housing assembly, for a detector of a sample separation apparatus for separating a fluidic sample, includes a lamp seat, a lamp insertable into the lamp seat, and a lamp cap mountable on the lamp seat and on the inserted lamp. The lamp seat, lamp and lamp cap are matched with respect to each other so that, by inserting the lamp into the lamp seat and by mounting the lamp cap on the lamp seat and on the inserted lamp, the lamp is axially and radially aligned and electrically and thermally coupled with the lamp seat and the lamp cap.