Hitrap mabselect sure 1 ml columns

4 Column PCC (4C‐PCC) was performed on a Semba Octave™ 10 BioSMB (Semba Biosciences, Madison, WI, USA) with five pumps attached to the four inlet ports. The inlet ports feed to the valve block with four HiTrap MabSelect SuRe™ 1 mL columns, with a bed height of 2.5 cm, attached (GE). The valve block is controlled by Semba™ Pro software and allows control of the inlets and outlets of each column, including running columns in series. A schematic is shown in Figure 3.
The buffers used for each inlet are shown in Table 4. The Elution outlet was fed directly to a Semba Octave™ 4XE UV–VIS Detector (Semba Biosciences) fitted with a preparative flow cell (0.3 mm). Fractionation details are provided in Single Cycle PCC runs and 100 Cycle Experiments sections, of the Materials and Methods, for single cycle and 100 cycle PCC runs, respectively.
Throughout the 4C‐PCC, each column is one of seven positions with its own combination of inlets and outlets (see Table 6). The feed and flowthrough steps feature two columns in series, with the outlet of the feed column used as the inlet for the flowthrough column. The wash/strip/equilibration outlet contains the post‐load wash, acid strip, or equilibration steps combined into a single outlet.

An ÄKTA™ Avant chromatography system (GE, Uppsala, Sweden) was used for all single column experiments. HiTrap MabSelect SuRe™ 1 mL columns (GE), with a bed height of 2.5 cm, were use. Separate columns were used for depth filter and FT‐AEX clarified feed materials. All samples were applied to the columns using the sample pump.