Multimodal Chromatography Media

Extracted from Multimodal Chromatography (PDF), GE Healthcare, 2013

GE Healthcare offers multimodal media in a wide range of formats to meet users’ needs at all stages of protein purification process development and manufacture. All of the multimodal chromatography media from GE Healthcare are BioProcess™ media. BioProcess media are developed and supported for production-scale chromatography. They are produced with validated methods and are tested to meet manufacturing requirements. Secure ordering and delivery routines give a reliable supply of media for production scale. Regulatory Support Files (RSF) are available to assist process validation and submissions to regulatory authorities. BioProcess media cover all purification steps from capture to polishing. The first multimodal medium introduced by GE Healthcare was Capto MMC, shortly followed by Capto adhere. As with all Capto media, the chosen ligands are coupled on a high-flow agarose base matrix, which gives improved pressure-flow properties compared with older media (Fig 3.1).

Comparison of the window of operation at large scale for different base matrices

Fig 3.1. Comparison of the window of operation (area below the curves) at large scale for different base matrices. Gray lines give the residence time in the column in minutes. The particle size of Capto MMC/adhere is 75 μm, and of Capto MMC/adhere ImpRes it is 40 μm. See Appendix 1 for characteristics of the various multimodal media from GE Healthcare.

The highly cross-linked agarose base matrix gives the media high chemical and physical stability. High flow velocities increase the productivity of large-scale bioprocessing operations and allow large volumes to be processed in one working shift. The multimodal ligands in Capto adhere and Capto MMC media were developed to offer novel selectivities compared with anion and cation exchangers, respectively. The multimodal ligands are now also available on the high-resolution base matrix Capto ImpRes (Fig 3.1), as Capto adhere ImpRes and Capto MMC ImpRes. GE Healthcare has also recently introduced Capto Core 700, a product that combines GF and multimodal anion exchange characteristics. Table 3.1 provides an overview of available multimodal chromatography products from GE Healthcare.

Table 3.1. Characteristics, benefits, and uses of multimodal chromatography media

Medium Structure Main functionalities Advantages Uses
Capto adhere Ligand:
N-benzyl methyl
ethanolamine
(see Fig 3.2)
Base matrix:
Capto
Electrostatic, interaction, hydrogen bonding, and hydrophobic interaction   High capacity and productivity
Removal of impurities to formulation levels in post-protein A purification
Wide operational window of pH and conductivity
Savings in time and operating costs with a two-step chromatographic process (see also Appendix 3, Use of multimodal media for MAb purification)
Intermediate purification and polishing of Mabs after capture on protein A.
Traditionally used in flowthrough mode.
Purification of other target proteins from capture to polishing steps.
Capto adhere ImpRes Ligand:
N-benzyl methyl
ethanolamine
(see Fig 3.2)
Base matrix:Capto ImpRes
 
See Capto adhere Same as Capto adhere but with higher resolution and lower elution volumes Efficient Mab polishing, removal of aggregates and HCP, and separations of charge variants. Polishing resulting in smaller elution volumes.

The properties of the small Capto ImpRes particle are best utilized in bind/elute mode.
Capto MMC Ligand:
N-benzoyl-homocysteine
(see Fig 3.6)
Base matrix:
Capto
Thiophilic interaction, hydrophobic interaction, hydrogen bonding, and electrostatic interaction Capto MMC gives high productivity and reduced cost with:
-High dynamic binding
-capacity (DBC) at high
-conductivity
-High volume throughput
-Different selectivity compared with traditional ion exchangers
Capture and intermediate purification of proteins from large feed volumes by packed bed chromatography. Purification can be performed at theconductivity of the feed material.  
Capto MMC ImpRes Ligand: N-benzoyl-homocysteine
(see Fig 3.6)
Base matrix:
Capto ImRes
See Capto MMC Same as Capto MMC but with higher resolution, lower elution volumes, and increased possibility to elute with salt only. Efficient Mab polishing, removal of aggregates and HCP, and separations of charge variants. Polishing resulting in smaller elution volumes.

The properties of the small Capto ImpRes particle are best utilized in bind/elute mode.
Capto Core 700 Ligand:
octylamine
(see Fig 3.12)
Base matrix:
high-flow agarose
GF, AIEX, and HIC Core bead technology with ligand-activated core and nonfunctionalized shell allows efficient capture of contaminants while target molecules are collected inflowthrough.   Significantly improved productivity compared with GF (100-fold)

Straightforward optimization and robust performance
Purification of viruses and other large target molecules
Custo Designed Media1 (CDM) Wide selection Wide selection Tailored to the user’s needs Various

 1 For challenging separation where standard multimodal media do not provide the desired results, CDM can provide libraries of additional multimodal anion or cation exchangers. The libraries are provided in 96-well microtiter plate format for rapid media screening. The multimodal cation and anion plates, respectively, contain 16 different multimodal media each.

Materials

     
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