Methods and systems are provided to improve consistency and robustness in chromatography, for example, in operations of multi-column chromatography systems.

A process for purifying a composition comprising water, a target substance, impurities and optionally cells, the process comprising the steps (A) and (B): (A) preparing a liquid feedstock by performing step (Ai) and/or (Aii) on the composition: (Ai) removing at least some of the cells from the composition; (Aii) concentrating the composition by removing water therefrom; and (B) passing the liquid feedstock through an apparatus comprising at least two processing units, each such unit producing a product stream containing purified target substance and optionally a waste stream comprising at least some of the impurities, wherein each unit comprises specified components (i) to (v). The units may be essentially the same except for a device they contain, leading to advantages in terms of simplicity, cost and ease of operation, lower risk of operator error, easier maintenance and lower inventory of spare parts.

A process for purifying a composition comprising water, a target substance, impurities and optionally cells, the process comprising the steps (A) and (B): (A) preparing a liquid feedstock by performing step (Ai) and/or (Aii) on the composition: (Ai) removing at least some of the cells from the composition; (Aii) concentrating the composition by removing water therefrom; and (B) passing the liquid feedstock through an apparatus comprising at least two processing units, each such unit producing a product stream containing purified target substance and optionally a waste stream comprising at least some of the impurities, wherein each unit comprises specified components (i) to (v). The units may be essentially the same except for a device they contain, leading to advantages in terms of simplicity, cost and ease of operation, lower risk of operator error, easier maintenance and lower inventory of spare parts.

The present disclosure provides, in some embodiments, devices, methods, and kits for purifying extracellular vesicles (EVs) using size exclusion chromatography in tandem with cation exchange chromatography, which can be referred to as dual-mode chromatography (DMC).

The present invention relates generally to processes for production of heavily glycosylated recombinant proteins (e.g., mucins and mucin-like proteins, such as lubricin), the processes comprising culturing mammalian cells capable of producing a glycoprotein in a liquid medium in a system comprising one or more bioreactors, concentrating and purifying and formulating the glycoprotein, the purification comprising one or more steps of chromatography, an endonuclease step, and at least one step of viral inactivation. In certain aspects the invention relates to pharmaceutical compositions comprising purified recombinant human lubiricin, and methods of treating a subject in need thereof.

In one aspect, a method for automated analysis of samples from a bioreactor is provided herein, the method including: drawing at least one sample from a bioreactor; pressurizing the drawn at least one sample into a sample flow; purifying at least one target protein in the sample flow using a first liquid chromatography apparatus to create a purified sample flow; splitting the purified sample flow into a purified sample fraction flow and an effluent flow; and, analyzing the at least one target protein in the purified sample fraction flow using a second liquid chromatography apparatus. Advantageously, the subject invention provides for an automated two-step liquid chromatography process utilizing first dimension liquid chromatography for purification and second dimension liquid chromatography for analysis.

A controller (50) of a liquid chromatograph (1) is configured to execute, as an analysis operation in an analysis unit (3), a sample injection step of bringing a high-pressure valve (10) into a loading state, sucking a sample from a tip of a needle (12) to hold a sample in a sampling channel (2), then connecting the sampling channel (2) to an injection port (16) and bringing the high-pressure valve (10) into an injecting state, and supplying a mobile phase from a liquid supplier (6), thereby injecting a sample held in the sampling channel (12) into an analysis channel (4), and an analysis step of separating components of a sample injected into the analysis channel (4) in a separation column (14) by bringing the high-pressure valve (10) in the loading state and supplying the mobile phase from the liquid supplier (6) after the sample injection step is ended. In a case where at least a predetermined condition is satisfied, after the analysis step is ended, the controller is configured to execute, as the analysis operation, a system cleaning step of cleaning a liquid flowing route from the sampling channel (2) to the analysis channel (4) by connecting the sampling channel (2) to the injection port (16) and bringing the high-pressure valve (10) into the injecting state, and supplying the mobile phase and/or a cleaning liquid from the liquid supplier (6).

A controller (50) of a liquid chromatograph (1) is configured to execute, as an analysis operation in an analysis unit (3), a sample injection step of bringing a high-pressure valve (10) into a loading state, sucking a sample from a tip of a needle (12) to hold a sample in a sampling channel (2), then connecting the sampling channel (2) to an injection port (16) and bringing the high-pressure valve (10) into an injecting state, and supplying a mobile phase from a liquid supplier (6), thereby injecting a sample held in the sampling channel (12) into an analysis channel (4), and an analysis step of separating components of a sample injected into the analysis channel (4) in a separation column (14) by bringing the high-pressure valve (10) in the loading state and supplying the mobile phase from the liquid supplier (6) after the sample injection step is ended. In a case where at least a predetermined condition is satisfied, after the analysis step is ended, the controller is configured to execute, as the analysis operation, a system cleaning step of cleaning a liquid flowing route from the sampling channel (2) to the analysis channel (4) by connecting the sampling channel (2) to the injection port (16) and bringing the high-pressure valve (10) into the injecting state, and supplying the mobile phase and/or a cleaning liquid from the liquid supplier (6).

Separation of materials is achieved using affinity binding and acoustophoretic techniques. A column provided with a fluid mixture of materials for separation and support structures may be used with acoustic waves to block flow of the support structures. The support structures can have an affinity for one or more materials in the fluid mixture. By blocking flow of the support structures, materials bound or adhered to the support structure are also blocked.

An ion exchange resin bag 5 includes a bag body 51 and a reinforcing body 52. The bag body 51 has a bottom surface portion 511 that is provided at an end portion opposite to an end portion where an opening is provided and forms a bottom surface of the bag body, and a side surface portion 512 that is connected to the bottom surface portion 511 and forms a side surface of the bag body 51. The reinforcing body 52 has a first reinforcing portion 521 that is fixed to a boundary portion of the bottom surface portion 511 and the side surface portion 512, and a second reinforcing portion 522 that is connected to the first reinforcing portion 521 and fixed to at least a part of the side surface portion 512 and extends from the first reinforcing portion 521 toward the opening.