Biopharmaceuticals

Biopharmaceuticals (Biological Source)

Biopharmaceutical manufacturing processes have high costs due to the multistep purification process, which is required to separate the many components generated during the cell culture step. The manufacturing process is roughly divided into upstream processes, including cell culture, and downstream processes, including purification. Purification using chromatography in the downstream process includes three steps: initial purification (capture), intermediate purification and final purification (polishing). In each step, the packing material (bulk medium) and chromatography device are chosen depending on the intended purpose.

Based on the interaction between the solid stationary phase and biomolecules, chromatographic techniques can be summarized into five classes: (i) affinity, (ii) ion-exchange, (iii) hydrophobic interactions, (iv) size exclusion and (v) mixed-mode chromatography.

Zeochem’s irregular shaped ZEOprep silica is a column packing material approved over decades for allowing highly reproducible results. Used primarily in upstream and downstream stages of biopharmaceutical processes, ZEOprep and ZEObead products offer high performance, high permeability and excellent selectivity, in addition to effective and reproducible separation.

Zeochem’s ZEOsphere DRP Mixed-Mode silica is an excellent choice for the purification of charged intermediates and as a polishing step. ZEOsphere DRP Mixed-Mode can substantially increase recovery/yield while decreasing organic solvent usage due to improved selectivity capabilities.


Applications

ZEOsphere silica can be used for developing a robust production process to purify APIs.

With over 25 years of expertise in the development of preparative Liquid Chromatography purification processes, ZEOsphere silica has proven to be a reliable partner in the delivery of high-quality, reproducible silica-based stationary phases for HPLC-based purification processes.

Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) is widely used in the biopharmaceutical industry. Analytical RP-HPLC is used for the release and characterization of raw materials, intermediates and APIs. Preparative HPLC is used for the commercial production of APIs that are not amenable to crystallization. The currently used stationary phases are not always suited for this task due to required cost reductions and the increasing complexity of APIs.

ZEOsphere DRP Mixed-Mode silica is an excellent answer for the purification of charged intermediates and as a polishing step for APIs. ZEOsphere DRP Mixed-Mode materials can substantially increase recovery/yield while decreasing purification costs through improved selectivity capabilities.

  • Amino acids
  • Biosimilars
  • Peptides
  • Proteins
  • Oligonucleotides
  • Vaccines

Custom Solutions

Preparative column chromatography (advanced normal phase and reversed phase chromatography) plays an important role in purifying valuable compounds used in research, pilot plant operations and production. Zeochem's ZEOsphere product line delivers high purity and high yield in an efficient manner.

ZEOsphere stationary phases will deliver reproducible results within narrow band ranges. This provides the highest degree of reliability for chromatographic separations and purifications.

Related Products

ZEOsphere silica is an excellent choice for the purification of (charged) intermediates and as a polishing step. ZEOsphere DRP Mixed-Mode can substantially increase recovery/yield while decreasing purification costs through improved selectivity capabilities.

Classical Reversed Phase – C18

ZEOsphere C18 phases are the most preferred as they offer an excellent range of hydrophobic separation power along with high surface area coverage.

Classical Reversed Phase – C4

ZEOsphere C4 is commonly used to separate large macromolecules such as proteins

Classical Reversed Phase – C8

An excellent range of hydrophobic separation power along with high surface area coverage, but when less retention compared to a C18 is needed.

Classical Reversed Phase – CN

Common applications based on Cyano phases include the separation of flavonoids, extraction of polar compounds from non-polar samples as well as analytes with a wide range of hydrophobicity.

Classical Reversed Phase – Phenyl and Derivatives

Classic reversed phase material based on 100Å and 120Å Ultra-pure silica with high surface area. ZEOsphere Phenyl as reverse phase or in HILIC mode shows alternative selectivity to C18 and C8 column

ZEObeads 50 C18 / 20-30µm

Spheroidal shape alkyl-bonded silica used in reversed-phase chromatography, preferably where superior flow performance and selectivity are needed.

ZEObeads 50 C8 / 20-30µm

ZEObeads 50 C8 / 20-30um is a spheroidal shaped silica with a reduced alkyl chain length, slightly less hydrophobic in reverse phase chromatography. The shorter alkyl chain length allows in some cases a better interaction with the target compound.

ZEObeads 50 Diol / 20–30µm

ZEObeads 50 NH2 / 20-30um is a spheroidal shaped amine bonded silica. It is used in normal phase chromatography for target compounds with more alkaline pH and as an excellent metal scavenger under conditions of superior flow performance.

ZEObeads 50 NH2 / 20-30µm

ZEObeads 50 NH2 / 20-30um is a spheroidal shaped amine bonded silica. It is used in normal phase chromatography for target compounds with more alkaline pH and as an excellent metal scavenger under conditions of superior flow performance.


Frequently Asked Questions

  • The larger the molecular weight of the target molecule to be separated the larger the pore size needs to be. The sample molecules need to be able to interact with the pores of the particle to create an efficient separation mechanism. If the pores are smaller than your target molecule it will not be retained and elute immediately.
  • The larger the average pore size of the material the smaller will be the effective surface area. If you have less surface area available, your separation is generally less efficient. It is important to choose the right pore size for your target molecule.
  • Size and distribution of the silica particles effect the packing efficiency of the columns and therefore process performance.
  • The higher the packing density, the higher the surface area, the more interaction between eluent and separation material resulting in higher efficiency separation. However, the higher the packing density, the smaller the mean particle size is and therefore the higher the backpressure will be.

Bonding different chemical groups onto the surface of bare silica results in functionalized silicas which give higherselectivity towards specific molecules.

  • Bare/Silica has a high polar surface, it is the most popular stationary phase, ideal for conventional applications (normal phase separations, non-polar target molecules, pharmaceuticals, natural products)
  • C4; suited to molecules with large hydrophobic regions, peptides, proteins. Usually combined with a large pore size for big biomolecules (>1000Da)
  • PHE/phenyl; moderately nonpolar for aromatic compounds, aflatoxins, caffeine, phenols.
  • C8; highly hydrophobic pesticides, peptides, drugs
  • C18; more apolar than C8 and in reversed phase chromatography higher retention than C8. Has a non-polar surface, ideal for pharmaceuticals, steroids, fatty acids, peptides, proteins, pesticides, PCB’s
  • Amino/NH2has a medium polar surface, ideal for carbohydrates and nitrogen containing heterocycles and amines. Scavenger applications.
  • Diolhas a lesspolar surface (than bare silica) and still many hydroxy-groups, ideal for lipids; Our best option for many SFC applications.
  • Cyano/CN unique selectivity for polar analytes with widely different chemistry eg. Cyclosporine, carbohydrates
  • Thiol/SH; Mainly used for scavenging
  • SCX/SO4:Ion Exchange for strong cation
  • SAX/TMA; Ion exchange for weak anion
  • NH2P; Ion exchange for scavenging
  • Morphology of the silica particles; meaning bead shape
  • Surface area: this is a function of particle- and pore size
  • Surface functionalisation
  • Particle size distribution in terms of range and actual distribution of particles within the range (D10/D50/D90)
  • Irregular silica beads to be used in low -medium pressure systems
  • Spheroidal Silica beads can be used in low to high pressure systems
  • Spherical silica beads primarily used for high-resolution high-pressure systems