Aldehyde activated SepFast

Amine activated SepFast

Carboxyl activated SepFast

CNBr-activated SepFast

Epoxy-activated SepFast

Hydrazide-activated SepFast

Irreversible Thiol-Coupling SepFast

NHS-activated SepFast

Aldehyde activated SepFast MAG

Amine activated SepFast MAG

Carboxyl activated SepFast MAG

CNBr activated SepFast MAG

Epoxy activated SepFast MAG

Hydrazide activated SepFast MAG

NHS activated SepFast MAG

Irreversible Thiol-Coupling SepFast MAG

Protein A & Alkaline-stable Protein A SepFast Range

Introduction

Protein A SepFast is a family of affinity chromatography media for the purification of immunoglobulins. Its purification power has been well documented in various antibody purification applications, such as isolation and purification of classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media.

• Protein A SepFast HighRes: 20 – 50 µm, good for small column run at shorter residence time (i.e. higher flowrate);
• Protein A SepFast HighRes Endure: 20 – 50 µm, more alkaline-stable than Protein A SepFast HighRes, can be cleaned with 0.5 M NaOH;
• Protein A SepFast: 50 – 150 µm, good for most applications from small scale to large scale purification requirements (i.e. low column pressure);
• Protein A SepFast Endure: 50 – 150 µm, more alkaline-stable than Protein A SepFast, can be cleaned with 0.5 M NaOH;
• Protein A SepFast Large Bead: 150 – 350 µm, particularly good for processing viscous or cell-containing feedstocks (i.e. high settling rate and extremely low column pressure);
• Protein A SepFast Large Bead Endure: 150 – 350 µm, more alkaline-stable than Protein A SepFast Large Bead, can be cleaned with 0.5 M NaOH;

Standard protein A molecule or more alkaline-stable protein A is immobilised to highly porous and highly cross-linked agarose base matrices. Agarose has long been used for chromatographic separations due to its excellent hydrophilic and low non-specific-binding nature. The particles have an open pore structure with excellent mass transfer properties to large protein molecules. The medium shows high mechanical rigidity, so it can be operated at moderate to high flow velocities with moderate pressure drop.

BioToolomics offers both loose media and pre-packed ready-to-use disposable columns for small scale sample preparation and large-scale bioprocessing use.

The range of Protein A SepFast media is developed and supported for production scale chromatography use. Regulatory Support File (RSF) is available to assist process validation and submissions to regulatory authorities.

Characteristics of Protein A SepFast Range & Protein A SepFast Endure Range

*Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)
** Please note that there might be considerable deviations in binding capacity for different immunoglobulins derived from the same species, even if they are of the same subclass.

Products

*Other column size available on request.

Ion-exchange (IEX) chromatography media is popularly employed to separate molecules, such as large proteins, small nucleotides and amino acids etc, based on their charge. The functional groups in IEX media have charges that bind molecules with opposite charges. Release of the bound molecules from the medium is achieved by displacement, via the application of an increasing concentration of a similarly charged molecule.

By design of the bead size, pore size, functional groups and the mechanical strength of individual base matrices, our IEX media can be used for capture of bulk product or impurities from large-volumes, as an intermediate purification step, or as a final step for high resolution purifications.

We offer an extensive range of ion exchange chromatography media (resins) in lab packs, bulk quantities and prepacked columns. Depending on the nature of your application, the property of your feedstock and the resolution you need, the general selection guide is shown along the product list below.

MabPolish® (Type I, Type II)

MabPolish is special mixed-mode chromatography media having strong binding to a broad range of host cell proteins and other impurities but little binding to antibodies (typically >90% yield). It is designed for process scale production of antibodies that requires regulatory support files.

MabPolish can be used in flow-through mode to remove impurities from monoclonal antibodies or other antibody materials, at various stages of the whole antibody purification process. Examples of the possible application scenario could be:

• Direct removal of impurities from crude clarified antibody materials
• After an antibody is first purified by affinity chromatography media (e.g. Protein A, Protein G or Protein L etc);
• After an antibody is first purified by ion-exchangers;
• After an antibody is first processed through precipitation

The base matrix is made of beaded agarose that has been highly cross-linked. The media is stable in most chemical conditions experienced in the bioprocessing industry.

Characteristics of MabPolish

Products

*Other column size available on request.

MabPolishTM DUO

MabPolish DUO is a class of novel resins with inert shell in the outer-layer of the bead (see the diagram below). The shell has size-exclusion effect that blocks molecules based on their molecular weights. Impurities smaller than a mAb can penetrate the shell and then be captured by the mixed-mode ligand inside the bead. It is a very gentle method with little loss of product.

The figure above shows the effectiveness of blocking IgG by the size-exclusion shell.

MabPolish DUO can be used in flow-through mode to remove impurities of less than 150 KDa from monoclonal antibodies or other antibody materials. Examples of the possible application scenario could be:

• After an antibody is first purified by affinity chromatography media (e.g. Protein A, Protein G or Protein L etc);
• After an antibody is first purified by ion-exchangers;
• After an antibody is first processed through precipitation;
• Direct removal of impurities from crude antibody materials
• After an antibody is modified (e.g. conjugated etc)

Characteristics of MabPolish DUO

Products

*Other column size available on request.

SepFast^TM DUO Range

SepFast DUO™ IEX is a range of unique shelled ion-exchange resins. Unlike ViralPolish® and MabPolish DUO, the core chemistries of the SepFast DUO™ media are classic ion-exchangers (Q or S).

SepFast DUO™ can be used for purifying viral materials in flow-through mode, impurities smaller than a virus will be adsorbed to the charged groups inside the bead whilst intact virus passes through the column. It is a very gentle method with little loss of product. Because the interior core has a standard ion-exchange ligand impurities can be eluted and the beads easily regeneration making this approach useful for large scale multi-use columns.

SepFast DUO 150 Q and SepFast DUO 150 S are particularly developed for purifying antibodies in flow-through mode. Host cell proteins (HCP) smaller than Ab molecules can be selectively removed.

SepFast DUO™ is available in four different outer shell pore sizes and two different inner core chemistries (see Table below)

Characteristics of SepFastTM DUO IEX Range

*Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

Products

*Other column size available on request.

Introduction

SepFast gel filtration media is specially designed for high resolution and high recovery fractionation of biological molecules based on their molecular weights. Gel filtration (or called size exclusion chromatography) is a proven technique, which is widely used for size-based molecular separations.

SepFast gel filtration media has a balanced design to offer high recovery and high selectivity according to individual applications. The core advantages are:

• High resolution
• High selectivity
• High recovery
• Highly scalable
• Excellent physical and mechanical stability
• Cost effective

The base matrix is made of agarose or a composite of polysaccharides that have been highly cross-linked. The media is very stable to most of the chemical conditions experienced in the bioprocessing industry.

The selection guide is listed as follows:

References Featuring Biotoolomics SepFast GF-HS-L Products:

de Cassan, S.C., Shakri, A.R., Llewellyn, D., Elias, S.C., Cho, J.S., Goodman, A.L., Jin, J., Douglas, A.D., Suwanarusk, R., Nosten, F.H. and Rénia, L., 2015. Preclinical assessment of viral vectored and protein vaccines targeting the Duffy-binding protein region II of Plasmodium vivax. Frontiers in immunology, 6, p.348

Jin, J., Tarrant, R.D., Bolam, E.J., Angell-Manning, P., Soegaard, M., Pattinson, D.J., Dulal, P., Silk, S.E., Marshall, J.M., Dabbs, R.A. and Nugent, F.L., 2018. Production, quality control, stability, and potency of cGMP-produced Plasmodium falciparum RH5. 1 protein vaccine expressed in Drosophila S2 cells. NPJ vaccines, 3(1), p.32

Characteristics of Gel Filtration Media

Products

*Other column size available on request.

Introduction

Based on the principle behind size exclusion chromatography, cross-linked neutral polysaccharide particles with very tight pores are packed into spin tubes for rapid desalting and / or buffer exchange.

Up to 24 samples (depending on the type of microcentrifuge) of 10 – 100 µl can be processed in one spin. It is a much faster and more efficient approach when compared to dialysis tubes or membrane ultrafiltration. The small porous particles provide huge surface area with very short diffusion distance, which means small molecules such as salts can be partitioned rapidly. In comparison, both dialysis tubes and membranes have very low surface areas. Dialysis is also incredibly time consuming, and membranes often suffer blockages resulting in a loss of valuable material.

Key benefits:

• Rapid desalting or buffer exchange
• Very little loss of target molecules (typically > 95% recovery)
• Most proteins (> 6,000 dalton) and DNAs (> 10 bp) can be desalted
• DNAse free
• Full operation manual supplied

SuperSpin desaltor is particularly useful for the following applications:

• Desalting of histidine-tagged proteins (e.g. imidazole and NaCl) recovered from IMAC SuperSpin
• Desalting of samples before loading to SDS-PAGE
• Desalting of samples before conducting other analysis
• Buffer exchange, for example, after low pH elution
• Desalting of DNAs

Technical data

Products

Introduction

Rapid concentration of samples up to ten times can be achieved in less than 4 minutes using the patented technology (patent pending).
Up to 24 samples (depending on the type of microcentrifuge) can be processed in one spin.

Dry microporous particles are packed into each spin tube for rapid removal of water and other small molecules.

It is a much faster and reliable approach when compared to ultrafiltration membranes (e.g. membrane blockage and severe loss of material) or vacuum evaporation (e.g. long processing time with special equipment).
Both proteins (> 6,000 dalton) and DNAs (> 10 bp) can be concentrated in a single spin.

The typical sample loading volume is 200 to 220 µl in each SuperSpin Concentrator.

Key benefits:

• Rapid sample concentration in very mild conditions in comparison to membrane ultrafiltration and vacuum evaporation methods
• Rapid concentration of DNAs in comparison to the ethanol or polymer precipitation methods
• Direct collection of the concentrated sample in the pass through, so little chance of contamination
• High recovery yield of target molecules (typically > 90%)
• Desalting can be achieved as well (if applicable)
• DNAse free
• Full operation manual supplied

SuperSpin Concentrator is particularly useful for the following applications:

• Concentration of small volume of protein samples
• Concentration of DNAs after Mini Preps
• Concentration of DNAs after cleaning up the PCR and restrictive digestion products etc

Technical data

Products

Introduction

Glutathione SepFast is an affinity chromatography medium used for the rapid one-step purification of Glutathione S-transferase (GST)-tagged proteins. Other Glutathione S-transferases and Glutathione-binding proteins can also be purified with this adsorbent.

Reduced Glutathione ligand is immobilised to the carefully designed porous support via very stable thiolether linkage.

GST-tagged proteins expressed in bacteria, yeasts, insects and mammalian cell cultures can be readily purified in a single purification step. GST tag can be cleaved in bound condition or in eluted condition by specific proteases.

Glutathione SepFast adsorbents are specifically designed and fabricated for the purification of GST-tagged proteins in batch (stirred tank), gravity flow or packed column modes. Glutathione SepFast is made of highly cross-linked agarose beads. Its carefully controlled pore structure allows fast access of affinity ligands by target protein molecules. Also, its high mechanical strength permits liquid to pass through gravity columns or packed columns at excellent flow rates. Clarified or unclarified cell lysates (for intracellular proteins) or culture broths (for extracellular proteins) could be directly processed with Glutathione SepFast. All these translate into high process flexibility and higher protein yield at shortened purification time.

Glutathione SepFast BG is supplied as loose resin or is supplied as pre-packed columns.

Characteristics of Glutathione SepFast resins and columns

Products

Introduction

Glutathione SepFast MAG is a magnetic affinity chromatography medium used for the rapid one-step purification of Glutathione S-transferase (GST)-tagged proteins with the aid of a magnet. Other Glutathione S-transferases and Glutathione-binding proteins can also be purified with this adsorbent.

Reduced Glutathione ligand is immobilised to the carefully designed magnetic support via very stable thiolether linkage.

GST-tagged proteins expressed in bacteria, yeasts, insects and mammalian cell cultures can be readily purified in a single purification step. After binding of target molecules, the magnetic resin can be readily isolated from the biological feedstocks with the aid of a magnet. The GST tag can be cleaved in bound condition or in eluted condition by specific proteases.

SepFast MAG is specially designed and fabricated for magnetic purification of proteins in batch mode (i.e. stirred tank mode). The base matrix is made of cross-linked agarose containing magnetic material. Therefore, the resin possesses magnetic properties. Removal of liquid after each step, such as binding, washing and elution can be readily done by fixing the resin with a magnet.

Its carefully controlled pore structure allows fast access of affinity ligands by target protein molecules. Also, its high mechanical strength permits harsher operational conditions. Clarified or unclarified cell lysates (for intracellular proteins) or culture broths (for extracellular proteins) could be directly processed with Glutathione SepFast MAG. All these translate into high process flexibility and higher protein yield at shortened purification time.

Most commercial magnetic devices can be used to handle this resin; no special magnet required. A magnetic stirring bar commonly available in laboratories can be used to drag down the magnetic particles.

Characteristics of Glutathione SepFast MAG

*Other particle size available on request.

Products

Introduction

Heparin SepFast is an affinity chromatography medium that is used for the purification of biomolecules that show affinity to heparin, such as antithrombin III, coagulation factors and other plasma proteins, DNA binding proteins, lipoproteins, protein synthesis factors, enzymes that act on nucleic acids, and steroid receptors.

The base matrix is made of cross-linked porous agarose. The particles have an open pore structure with good mass transfer properties to large protein molecules. Heparin is immobilized to the base matrix by reducing amination method.

The medium shows high mechanical rigidity, so it can be operated at high flow velocities with moderate pressure drops. The medium is compatible with most of the chemical reagents commonly used in biological systems.

Characteristics of Heparin SepFast Media

Products

SepFast Butyl Products

SepFast Phenyl Products

SepFast Octyl Products

Note: HIC media with lower or higher ligand density can be custom-made to increase recovery or binding capacity. Please contact us for more information.

Ion-exchange (IEX) chromatography media is popularly employed to separate molecules, such as large proteins, small nucleotides and amino acids etc, based on their charge. The functional groups in IEX media have charges that bind molecules with opposite charges. Release of the bound molecules from the medium is achieved by displacement, via the application of an increasing concentration of a similarly charged molecule.

By design of the bead size, pore size, functional groups and the mechanical strength of individual base matrices, our IEX media can be used for capture of bulk product or impurities from large-volumes, as an intermediate purification step, or as a final step for high resolution purifications.

We offer an extensive range of ion exchange chromatography media (resins) in lab packs, bulk quantities and prepacked columns. Depending on the nature of your application, the property of your feedstock and the resolution you need, the general selection guide is shown along the product list below.

Introduction

IMAC SepFast is designed for the purification of histidine-tagged proteins in batch, gravity flow, packed bed, or 96-well microplate operations.

Key benefits:

• Low cost due to the novel manufacturing processes
• High binding capacity (could be over 40 mg/ml)
• Fast and easy protein purification from unclarified or clarified samples
• Choice of three pre-charged metal ions
• Very low metal leakage
• Full protein purification manual supplied

Its carefully controlled pore structure allows fast access of the target protein to the immobilised metal ligand. Also, its high mechanical strength permits liquid to pass through a column at excellent flow rates. Clarified or unclarified cell lysates (for intracellular proteins) or culture broths (for extracellular proteins) can be directly processed with IMAC SepFast. All these translate into high process flexibility and higher protein yield at a shortened purification time.

IMAC SepFast or its three metal charged formats i.e. Ni SepFast, Co SepFast and Zn SepFast, are highly stable and compatible with a wide range of chemicals (e.g. detergents, denaturing reagents and reducing reagents etc) commonly used in protein purification processes, which means that more flexible operations can be developed for the best performance.

Product Characteristics

*Test with nickel ion charged; **Tested with the absence of metal ions.

Products

Introduction

Ni SepFast resin is packed into our robust and durable chromatography columns for convenient use. The column ends have 1/16″ fittings that are compatible with common chromatography systems (such as AKTA). The column body is robust enough for use in moderate pressure operations. The column has been well designed to minimise the dead volume and gives an excellent flow distribution across the bed.
The 1 ml column is made of polypropylene. The column body for the 5 ml column is made of acrylic, which gives a clear view of the packed resin.

Key benefits:

• Low cost
• High protein binding capacity with high purity
• Ready to use
• Robust and durable for re-use or longer storage
• Full operational manual supplied

We can also pack other type of chromatography resins that you choose into our columns with a reduced cost. Larger column sizes are also available. See our Column Packing Service for more information.

For large quantities of columns, please contact us directly and we will offer a very good price structure to satisfy your applications.

Characteristics of Pre-packed Ni SepFast Columns

*Tested with nickel ion charged

Products

Introduction

IMAC SuperSpin is supplied as a low cost disposable spin-column that allows rapid purification and screening of histidine-tagged proteins at a low cost. The spin tube is filled with novel metal immobilised chromatography resin of small particles (20 – 50 µm). It gives fast mass transfer rate of target proteins, so decent amounts of protein can be captured in a short solid / liquid contact time. Three types of spin tubes are available; Ni SuperSpin, Co SuperSpin and Zn SuperSpin, for fast screening of the best immobilised metal ion to a given target protein. It is a particularly powerful tool in applications such as small-scale purification, high-throughput screening, purification process optimisation etc.

Key benefits:

• Fast purification of many protein samples with a standard microcentrifuge
• High protein binding capacity
• Very low cost in comparison to similar products in the marketplace
• Optimisation of the binding, washing and elution conditions (e.g. imidazole concentration, pH, salt concentration, detergents etc) in one or two sets of experiments in short periods and at very low cost
• Direct processing of clarified or unclarified protein sample
• Full operational manual supplied

The fine IMAC particles are very stable and compatible with a variety of chemical reagents (e.g. denaturing reagents and reducing reagents etc).

Characteristics of Ni SuperSpin / Co SuperSpin / Zn SuperSpin

*Tested with nickel ion charged

Products

Introduction

Magnetic IMAC (immobilised metal affinity chromatography) resin is typically useful in purifying small quantity low concentration histidine-tagged proteins. For example, when 10 – 50 µl of resin is required for protein purification, it is difficult to handle such small quantities in columns. Magnetic IMAC resins can be easily processed with the aid of a magnet. The liquid / solid separation required in each purification step (e.g. equilibration, binding, washing and elution) is readily achieved with the aid of a magnet (e.g. magnetic stirring bar or other commercial magnetic devices).

Key benefits:

• Tailored base matrix design for batch magnetic purification of proteins
• High binding capacity (could be over 40 mg/ml)
• Simple and convenient operation, no chromatography training required
• Flexible operational conditions
• Low cost
• A choice of three types of immobilised metal ions available
• Very low metal leakage
• Full operation manual (including purification conditions) supplied

Product Characteristics

*Tested with nickel ion charged; **Tested with the absence of metal ion

Products

Note: No magnet is supplied with the above products. Customers need to source the magnet themselves.

Introduction

Glutathione SepFast MAG is a magnetic affinity chromatography medium used for the rapid one-step purification of Glutathione S-transferase (GST)-tagged proteins with the aid of a magnet. Other Glutathione S-transferases and Glutathione-binding proteins can also be purified with this adsorbent.

Reduced Glutathione ligand is immobilised to the carefully designed magnetic support via very stable thiolether linkage.

GST-tagged proteins expressed in bacteria, yeasts, insects and mammalian cell cultures can be readily purified in a single purification step. After binding of target molecules, the magnetic resin can be readily isolated from the biological feedstocks with the aid of a magnet. The GST tag can be cleaved in bound condition or in eluted condition by specific proteases.

SepFast MAG is specially designed and fabricated for magnetic purification of proteins in batch mode (i.e. stirred tank mode). The base matrix is made of cross-linked agarose containing magnetic material. Therefore, the resin possesses magnetic properties. Removal of liquid after each step, such as binding, washing and elution can be readily done by fixing the resin with a magnet.

Its carefully controlled pore structure allows fast access of affinity ligands by target protein molecules. Also, its high mechanical strength permits harsher operational conditions. Clarified or unclarified cell lysates (for intracellular proteins) or culture broths (for extracellular proteins) could be directly processed with Glutathione SepFast MAG. All these translate into high process flexibility and higher protein yield at shortened purification time.

Most commercial magnetic devices can be used to handle this resin; no special magnet required. A magnetic stirring bar commonly available in laboratories can be used to drag down the magnetic particles.

Characteristics of Glutathione SepFast MAG

*Other particle size available on request.

Products

Introduction

Magnetic IMAC (immobilised metal affinity chromatography) resin is typically useful in purifying small quantity low concentration histidine-tagged proteins. For example, when 10 – 50 µl of resin is required for protein purification, it is difficult to handle such small quantities in columns. Magnetic IMAC resins can be easily processed with the aid of a magnet. The liquid / solid separation required in each purification step (e.g. equilibration, binding, washing and elution) is readily achieved with the aid of a magnet (e.g. magnetic stirring bar or other commercial magnetic devices).

Key benefits:

• Tailored base matrix design for batch magnetic purification of proteins
• High binding capacity (could be over 40 mg/ml)
• Simple and convenient operation, no chromatography training required
• Flexible operational conditions
• Low cost
• A choice of three types of immobilised metal ions available
• Very low metal leakage
• Full operation manual (including purification conditions) supplied

Product Characteristics

*Tested with nickel ion charged; **Tested with the absence of metal ion

Products

Note: No magnet is supplied with the above products. Customers need to source the magnet themselves.

Introduction

Protein A SepFast MAG and Protein G SepFast are magnetic beads designed for coupling of antibodies enabling enrichment of target protein for further downstream analyses such as mass spectrometry (MS and LC-MS) and electrophoresis techniques. The base matrix is made of cross-linked agarose encapsulating magnetic material. Therefore, the resin possesses magnetic properties. Removal of liquid after each step such as binding, washing and elution can be readily done by fixing the resin with a magnet.

Our product can be used with any magnetic device commonly available in the market place or magnets of any shape.

Protein A and Protein G binds to the Fc region of immunoglobulins. The binding is highly specific so high purity can be achieved in a single step. The purification power of Protein-A ligand and Protein G ligand has been well documented in various antibody purification applications, such as isolation and purification of classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media. In general, protein G shows affinity to more classes of immunoglobulins than protein A.

Characteristics of Protein A SepFast MAG & Protein G SepFast MAG

*Tested with nickel ion charged

Products

Introduction

Magnetic SepFast™ Supor Q (DEAE, S, CM) is a group of special ion-exchange media with magnetic property. The working medium possesses a combination of small pores (50-100nm) and large pores (micro level). It shows fast accessibility to both small and large molecules (e.g. plasmid and virus). Therefore, it has comparable binding capacity to those of membrane or monolith product in respect of DNA or virus. However, Magnetic SepFast™ Supor media shows much higher binding capacity to protein molecules than the membrane or monolith type of products.

The base matrix is made of a composite of magnetic polysaccharides that have been well cross-linked. The media is stable to most of the chemical conditions experienced in the bioprocessing industry. The magnetic Supor media is particularly useful in the high-through-put screening of phage libraries or high-through-put purification of viruses or VLPs etc.

*Other volume sizes are available on request.

SepFast MM AH-1 is a mixed mode chromatography medium that is supplied in a ready-to-use disposable format. The ligand contains a combination of anionic and hydrophobic groups. The product shows good binding capacity to molecules rich in hydrophobic zones.

The base matrix is made of cross-linked porous polysaccharide. The particle has open pore structure with good mass transfer property. The particle size is 50 – 150µm. The medium is compatible with most of the chemical reagents commonly used in biological systems.

The medium is stored in 20% industrial ethanol at 4-30oC. Before a chromatography run, equilibrate the column with working buffer until the effluent shows stable conductivity and pH value.

The operating pressure is 4 bar or less.

SepFast MM AH-1 can be used for the capturing of proteins in a binding-elution mode. It can also be used in a flow-through mode to remove the impurities. We recommend scouting for the optimal binding, washing and elution conditions such as pH and conductivity.

Products

*Other volume sizes are available on request.

Introduction

BioToolomics developed ViralPolish® chromatography media specifically for virus purification; Comprising dual layer agarose beads with an inert external shell with tightly controlled pore size and internal polyfunctional ligands for rapid high capacity binding of impurities.

This media is ideal for rapid clean-up, either as a first step or as a second step, to purify virus particles.

Large particles, such as viruses, are excluded from the beads. The beads can be packed in a column where the virus will pass through the column bed and collected in the flow through fraction, whilst impurities will be captured within the beads. This affords a very gentle purification process resulting in efficient clean up and high recovery of active virus particles.

The beads can also be used in batch mode; when added to a flask of virus containing media the beads will selectively allow small molecules to enter and be retained, whilst the virus is excluded and remains in the external media.

A typical purification strategy for virus would be to use ViralPolish® as an initial step, to clean up the sample, prior to binding and elution, typically from an anion exchange media designed for virus purification such as SepFast Supor Q.

Alternatively, very dilute samples can be purified by first passing through a SepFast Supor Q anion exchange column to concentrate the sample followed by passing through ViralPolish column to remove residual impurities.

ViralPolish® is available in three different outer shell pore sizes, and two different impurity binding chemistries (see table below).

Note: other mixed-mode ligands are available on request.

Characteristics of ViralPolish® Range

*Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

The pore size distribution of the outer layer is critical to prevent molecules of a given molecular weight from binding to exposed ‘active core’. ViralPolish® has tightly controlled pores giving best in class exclusion of virus and large molecules, resulting in significantly higher recovery of active viral particles.

Products

*Other column size available on request.

Introduction

SepFast DUO™ IEX is a range of unique shelled ion-exchange resins. Unlike ViralPolish® and MabPolish DUO, the core chemistries of the SepFast DUO™ media are classic ion-exchangers (Q or S).

SepFast DUO™ can be used for purifying viral materials in flow-through mode, impurities smaller than a virus will be adsorbed to the charged groups inside the bead whilst intact virus passes through the column. It is a very gentle method with little loss of product. Because the interior core has a standard ion-exchange ligand impurities can be eluted and the beads easily regeneration making this approach useful for large scale multi-use columns.

SepFast DUO 150 Q and SepFast DUO 150 S are particularly developed for purifying antibodies in flow-through mode. Host cell proteins (HCP) smaller than Ab molecules can be selectively removed.

SepFast DUO™ is available in four different outer shell pore sizes and two different inner core chemistries (see Table below)

Characteristics of SepFastTM DUO IEX Range

*Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

*Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

The pore size distribution of the outer layer is critical to prevent molecules of a given molecular weight from binding to exposed ‘active core’. ViralPolish® has tightly controlled pores giving best in class exclusion of virus and large molecules, resulting in significantly higher recovery of active viral particles.

Products

*Other column size available on request.

Introduction

SepFastTM Supor Q (DEAE, S, CM) is a strong anion exchange chromatography product in a ready-to-use disposable format. The working medium possesses a combination of small pores (50-100nm) and large pores (micro level). It shows fast accessibility to both small and large molecules (e.g. plasmid and virus). Therefore, it has comparable binding capacity to those of membrane or monolith product in respect of DNA or virus. However, SepFastTM Supor media shows much higher binding capacity to protein molecules than the membrane or monolith type of products.

The base matrix is made of a composite of polysaccharides that have been well cross-linked. The medium is stable to most of the chemical conditions experienced in the bioprocessing industry.

It shows excellent binding capacity to both protein and macromolecule e.g. DNAs (see the Table 1) at high flowrate. Certainly the capacity is further increased at lower flowrates.

Table 1: A comparison of the dynamic binding capacity (DBC at 10% breakthrough) of different products (1 cm bed height):

SepFast^TM Supor IEX media can be used in the final polishing step (shallow bed format) or be used for the binding-elution of both small (proteins) and large molecules (e.g. plasmid and virus) at deep bed format. Please contact us for further information.

It can be operated at both high flowrate or low-to-moderate flowrate. The column is compatible to most of the liquid-handling equipment. The operational pressure should not exceed 4 bars.

The medium was stored in 20% denatured ethanol. Before a chromatography run, equilibrate the column with working buffer until the effluent shows stable conductivity and pH value.

We recommend scouting for optimal binding pH and for optimal ionic strength.

SepFast^TM Supor media has a protein binding capacity (tested with BSA) virtually independent of the flowrate. We recommend to pay special attention to optimising elution conditions to avoid tailing in the elution step.

References Featuring Biotoolomics SepFast Supor Q or SepFast Supor DEAE Products:

Lim, S.L et. al. 2018. Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography. Process Biochemistry, 69, pp.208-215.

Lim, S.L.et.al.  2015. Direct purification of hepatitis B core antigen Y132A dimer using packed bed anion exchange chromatography. In Asia Pacific Confederation of Chemical Engineering Congress 2015: APCChE 2015, incorporating CHEMECA 2015 (p. 1370). Engineers Australia.

Yap, C.F. et. al. 2013. Purification of long helical capsid of newcastle disease virus from Escherichia coli using anion exchange chromatography. Biotechnology progress, 29(2), pp.564-567.

Monjezi, R. et. al. 2010. Purification of bacteriophage M13 by anion exchange chromatography. Journal of Chromatography B, 878(21), pp.1855-1859.

Products

*Other volume size available on request.

For predictable screening and scaling-up of a chromatography purification step, reliable and reproducible column packing is very critical. We have extensive experience in handling a broad range of chromatography media, from soft to rigid, from small particles to large particles, from light particles to dense particles and from polymer-based particles to inorganic-based particles. The resin can be of any type you choose.

We can pack high performance and robust disposable chromatography columns from small to large (e.g. 1μl to 10 litres), for cGMP or non-cGMP applications, using our proprietary column design. The bed height can be anywhere between 0 cm and 60 cm.

Our range of chromatography columns has the following key features and advantages:

• Fully scalable column design for bioprocessing applications
• Low cost
• Resin compressibility taken into account
• Minimum dead volume for high resolution applications
• Proven flow distribution mechanism
• Disposable
• Tight packing of any type of chromatography media (>20 µm)
• Features standard connections, which are compatible with most common chromatography instruments such as AKTA etc.
• Easy to store
• Long life time

For each packed column we will test HETP, Peak Asymmetry and Flow Properties (if required). For cGMP grade columns, in addition to the above tests, we will also test Endotoxin and Bioburden levels.

We can pack a column according to your protocol or develop the best predictable packing process for you. According to your special applications, we can also tailor our column design to suit your needs.

The available column sizes and their characteristics can be seen in the tables below.

Please contact us to discuss your requirements in details.

We offer the following packing services for medium to high throughput purification applications:

We offer the following packing services for process development and manufacture applications:

Note:

Chemical compatibility: All products are compatible with most commonly used aqueous buffers. For special chemical or certain organic solvent compatibility please contact us.

*We could make customised columns of these diameters for cGMP use, please contact us for more information.

If you can’t find the suitable commercial media or the commercial media couldn’t match your purification needs, why not consider the option of customised media?

BioToolomics has extensive experience in fabrication of chromatography resins, in particular the polysaccharide-based media such as agarose. According to your application, we can design the right base matrix to match with the right ligand.

The key factors to customise a chromatography medium include: base matrix material, porosity, pore size, backbone chemistry, particle size and size distribution, particle density, cross-linking, particle rigidity, type of ligand, cost and availability of a ligand, stability of the ligand before and after the immobilisation, immobilisation chemistry, orientation of the ligand, ligand functionality and accessibility post the immobilisation, ligand density and ligand leakage etc.

We have the capability to make customised media in volumes that range from a few millilitres to tens of litres. So no matter your requirement is small or large, please contact us and we will find a solution for you.

Introduction

Biotoolomics offers scalable chromatography columns for fast medium screening, process development and scaling-up applications, or general laboratory purifications. The empty column sizes range from 1ml to 40ml, and packed column sizes range from 1ml to 10 litres.

Our range of chromatography columns has the following key features and advantages:

• Fully scalable column design for bioprocessing applications
• Low cost
• Resin compressibility taken into account
• Minimum dead volume for high resolution applications
• Proven flow distribution mechanism
• Disposable
• Easy to pack
• Tight packing of any type of chromatography media (>20 µm)
• Features standard connections, which are compatible with most common chromatography instruments such as AKTA etc.
• Easy to store
• Long life time

After the chosen resin is packed in the column, the column can be readily stored and re-used for many times.

Characteristics of Empty Columns

For detailed information about individual columns please click the relevant column size below:

• 1 ml, 0.67 ml, 0.33 ml
• 5 ml, 10 ml
• 20 ml, 30 ml, 40 ml

For our full range of column sizes please click column packing service.

*For columns of other volumes or higher pressure ratings, please contact us for a quotation.

**The minimum order is 5 columns. If large quantity is required, please contact us for a quotation. BioToolomics also provides service to pack customers’ media of their choice.

For predictable screening and scaling-up of a chromatography purification step, reliable and reproducible column packing is very critical. We have extensive experience in handling a broad range of chromatography media, from soft to rigid, from small particles to large particles, from light particles to dense particles and from polymer-based particles to inorganic-based particles. The resin can be of any type you choose.

We can pack high performance and robust disposable chromatography columns from small to large (e.g. 1μl to 10 litres), for cGMP or non-cGMP applications, using our proprietary column design. The bed height can be anywhere between 0 cm and 60 cm.

Our range of chromatography columns has the following key features and advantages:

• Fully scalable column design for bioprocessing applications
• Low cost
• Resin compressibility taken into account
• Minimum dead volume for high resolution applications
• Proven flow distribution mechanism
• Disposable
• Tight packing of any type of chromatography media (>20 µm)
• Features standard connections, which are compatible with most common chromatography instruments such as AKTA etc.
• Easy to store
• Long life time

For each packed column we will test HETP, Peak Asymmetry and Flow Properties (if required). For cGMP grade columns, in addition to the above tests, we will also test Endotoxin and Bioburden levels.

We can pack a column according to your protocol or develop the best predictable packing process for you. According to your special applications, we can also tailor our column design to suit your needs.

The available column sizes and their characteristics can be seen in the tables below.

Please contact us to discuss your requirements in details.

We offer the following packing services for medium to high throughput purification applications:

Products

We offer the following packing services for process development and manufacture applications:

Note:

Chemical compatibility: All products are compatible with most commonly used aqueous buffers. For special chemical or certain organic solvent compatibility please contact us.

*We could make customised columns of these diameters for cGMP use, please contact us for more information.