A preparative liquid chromatograph includes a liquid feeding pump (2) that feeds a mobile phase, an injector (4) that injects a sample into the mobile phase at a downstream of the liquid feeding pump (2), a separation column (6) for separating components in the sample injected into the mobile phase by the injector (4) at a downstream of the injector (4), and an eluate fractionator (8) configured to divide a flow of the eluate from the separation column (6) into a flow of a minute flow rate and another flow at a downstream of the separation column (6) and to extract at least a part of an eluate that forms the flow of a minute flow rate into an fractionation container (22).

The invention is a method for liquid, gaseous or supercritical phase chromatography which involves circulating, on a chromatograph (6) containing a stationary phase, a load (1) comprising components to be separated entrained by a carrier fluid (2), said method being characterized in that it involves: (a) obtaining, at the outlet of the chromatograph, a plurality of chromatographic fractions (3, 4) comprising at least one component of the load and the carrier fluid in a first fluid phase, (b) imposing a change of state on at least one of said chromatographic fractions (3, 4) so as to obtain at least one fraction of purified carrier fluid in a second fluid phase different from the first fluid phase by separating said carrier fluid from the component of the load, (c) imposing a change of state in a reverse direction to that of step (b) on at least one fraction of purified carrier fluid obtained in step (b) so as to obtain at least one fraction of purified carrier fluid in a third fluid phase different to the second fluid phase, and in that it involves coupling the change-of-state energies from the first fluid phase to the second fluid phase and from the second fluid phase to the third fluid phase of the same or of another fraction of purified carrier fluid, said coupling comprising a transfer of heat using a heat pump.

The invention is a method for liquid, gaseous or supercritical phase chromatography which involves circulating, on a chromatograph (6) containing a stationary phase, a load (1) comprising components to be separated entrained by a carrier fluid (2), said method being characterized in that it involves: (a) obtaining, at the outlet of the chromatograph, a plurality of chromatographic fractions (3, 4) comprising at least one component of the load and the carrier fluid in a first fluid phase, (b) imposing a change of state on at least one of said chromatographic fractions (3, 4) so as to obtain at least one fraction of purified carrier fluid in a second fluid phase different from the first fluid phase by separating said carrier fluid from the component of the load, (c) imposing a change of state in a reverse direction to that of step (b) on at least one fraction of purified carrier fluid obtained in step (b) so as to obtain at least one fraction of purified carrier fluid in a third fluid phase different to the second fluid phase, and in that it involves coupling the change-of-state energies from the first fluid phase to the second fluid phase and from the second fluid phase to the third fluid phase of the same or of another fraction of purified carrier fluid, said coupling comprising a transfer of heat using a heat pump.

The present invention provides methods to improve the sensitivity of detecting glycosylamines released from glycoconjugates, such as glycoproteins or glycopeptides, by enzymatic digestion when labeling them with amine-reactive dyes.

In some aspects, the present disclosure pertains to sample treatment methods that comprise: contacting an acidic elution solution that is free of primary amine, secondary amine and thiol groups with a sorbent having bound target antibody and separating the elution solution from the sorbent, thereby releasing bound target antibody from the sorbent and forming a first collection fraction that comprises the elution solution and released target antibody; contacting the sorbent with a neutralization buffer solution that is free of primary amine, secondary amine and thiol groups and separating the neutralization buffer solution from the sorbent, thereby forming a second collection fraction that comprises the neutralization buffer solution; and forming a neutralized solution that comprises the first collection fraction and the second collection fraction. In other aspects, the present disclosure pertains to kits for performing such sample treatment methods.

Presteralized manifolds are provided which are designed for sterile packaging and single-use approaches. Disposable tubing and flexible-wall containers are assembled via aseptic connectors. These manifolds are adapted to interact with other equipment which can be operated by a controller which provides automated and accurate delivery of biotechnology fluid. The manifold also can be used in conjunction with one or more sensors such as pressure and conductivity sensors that interact with the controller or are connected to a separate user interface. An aseptic environment obtains avoiding or reducing cleaning and quality assurance procedures.

Presteralized manifolds are provided which are designed for sterile packaging and single-use approaches. Disposable tubing and flexible-wall containers are assembled via aseptic connectors. These manifolds are adapted to interact with other equipment which can be operated by a controller which provides automated and accurate delivery of biotechnology fluid. The manifold also can be used in conjunction with one or more sensors such as pressure and conductivity sensors that interact with the controller or are connected to a separate user interface. An aseptic environment obtains avoiding or reducing cleaning and quality assurance procedures.

A permeative amine/acid introduction device (PAID) is placed after a conventional KOH eluent suppressed conductometric anion chromatography (SCAC) system. The PAID converts the suppressed eluites from the acid form to the corresponding salt. For example, when the analytes are acids, they are converted to the corresponding ammonium salt (NR.sub.2H+HX.fwdarw.NR.sub.2H.sub.2.sup.++X.sup.−) and allows very weak acids HX (pK.sub.a≥7.0) that cannot normally be detected by SCAC to be measured by a second conductivity detector following the PAID. Permeative reagent introduction is dilutionless, can be operated without pumps and provides good mixing with low band dispersion (as small as 30 μL). An exemplary amine is diethylamine (DEA), which was chosen as the amine source due to its low pK.sub.b value (pK.sub.b 3.0), high vapor pressure, and low toxicity and low odor.

A permeative amine/acid introduction device (PAID) is placed after a conventional KOH eluent suppressed conductometric anion chromatography (SCAC) system. The PAID converts the suppressed eluites from the acid form to the corresponding salt. For example, when the analytes are acids, they are converted to the corresponding ammonium salt (NR.sub.2H+HX.fwdarw.NR.sub.2H.sub.2.sup.++X.sup.−) and allows very weak acids HX (pK.sub.a≥7.0) that cannot normally be detected by SCAC to be measured by a second conductivity detector following the PAID. Permeative reagent introduction is dilutionless, can be operated without pumps and provides good mixing with low band dispersion (as small as 30 μL). An exemplary amine is diethylamine (DEA), which was chosen as the amine source due to its low pK.sub.b value (pK.sub.b 3.0), high vapor pressure, and low toxicity and low odor.

The present application provides methods and systems for viral clearance for purifying an antibody from a sample comprising one or more impurities including viral particles. The method is conducted in a system which includes a hydrophobic interaction chromatography (HIC) column and a virus retentive filtration (VRF) system. The HIC column and the VRF system are connected inline in a continuous processing system, and the VRF system comprises at least two filter trains in parallel.