75um Agarose Gel Filter Packing Hard GF 3.1 HardGF 3.1

75 μm High-Rigidity Agarose Gel Filtration Media HardGF 3.1

SHBC HardGF 3.1 is a 75 μm high-rigidity agarose gel filtration medium manufactured from 3.1% agarose through emulsification, washing and multiple crosslinking processes.

The resulting crosslinked agarose microspheres are designed for size exclusion chromatography (SEC), also known as gel filtration chromatography, for separation and purification of proteins, biomolecules and macromolecular samples.

HardGF 3.1 combines a hydrophilic agarose matrix with enhanced bead rigidity for laboratory purification, process development and scalable chromatography.

HardGF 3.1 Technical Specifications

What Is HardGF 3.1 Agarose Gel Filtration Media?

HardGF 3.1 is a porous agarose-based chromatography medium designed for size-based biomolecule separation.

It is prepared from 3.1% agarose through:

Agarose Preparation → Emulsification → Microsphere Formation → Washing → Multiple Crosslinking → High-Rigidity Agarose Beads

Multiple crosslinking strengthens the agarose network while retaining the hydrophilic porous structure required for gel filtration chromatography.

Commercial SEC products also use crosslinked agarose matrices to improve physical stability and packed-bed performance. Cytiva's Sepharose 4 Fast Flow, for example, uses a crosslinked 4% agarose matrix specifically designed for process-scale size exclusion chromatography.

High-Rigidity Crosslinked Agarose Structure

Traditional soft agarose gels may deform under increasing flow or bed pressure.

HardGF 3.1 uses repeated crosslinking to create a stronger three-dimensional agarose network.

Key structural features include:

• 3.1% agarose matrix

• Multiple crosslinking

• High bead rigidity

• Hydrophilic surface

• Porous internal structure

• Spherical particle format

• 75 μm nominal particle size

• Suitable for packed-bed chromatography

Crosslinking is also used in established commercial agarose SEC media to increase matrix stability. Cytiva describes crosslinked agarose as providing improved physical stability and process performance.

How Size Exclusion Chromatography Works

HardGF 3.1 separates molecules primarily according to their effective hydrodynamic size.

Larger Molecules

Large molecules have limited access to the internal pores of the agarose beads.

They therefore travel through the column more directly and generally elute earlier.

Smaller Molecules

Smaller molecules can enter more of the internal pore volume.

They follow a longer pathway through the packed bed and generally elute later.

The separation requires no specific binding between the analyte and resin.

Key Advantages of HardGF 3.1

• 75 μm nominal particle size

• 3.1% agarose matrix

• Multiple-crosslinked structure

• High-rigidity agarose microspheres

• Hydrophilic chromatography matrix

• Size-based separation mechanism

• Low dependence on specific ligand interactions

• Suitable for packed-column chromatography

• Suitable for protein and biomolecule purification

• Suitable for laboratory and process development

• Bulk production available

Why Choose 75 μm Agarose Beads?

Particle size is an important factor affecting column efficiency, pressure and flow characteristics.

The 75 μm particle format provides a practical balance for preparative chromatography applications.

Potential benefits include:

• Efficient packed-bed formation

• Good mass transfer

• Suitable chromatographic resolution

• Practical process flow characteristics

• Convenient column packing

• Suitable for preparative purification

• Suitable for scale-up studies

Bio-Rad currently offers agarose SEC media in 37–75 μm and 75–150 μm particle ranges, showing that approximately 75 μm is an established particle-size class for preparative agarose gel filtration.

Protein and Biomolecule Separation

HardGF 3.1 can be evaluated for size-based separation of:

• Proteins

• Recombinant proteins

• Antibodies

• Protein complexes

• Protein aggregates

• Enzymes

• Polysaccharides

• Nucleic acid-related macromolecules

• Other biological macromolecules

The actual suitable molecular-weight separation range should be determined using HardGF 3.1 product-specific chromatography data.

Antibody and Protein Aggregate Purification

Size exclusion chromatography is frequently used to separate monomeric proteins from higher-molecular-weight aggregates and lower-molecular-weight impurities.

Potential HardGF 3.1 applications include:

• Antibody polishing

• Protein aggregate removal

• Protein monomer separation

• Protein complex analysis

• Macromolecule fractionation

• Sample cleanup

Bio-Rad specifically positions its agarose SEC media for purification of antibodies and aggregates.

Large Biomolecule and Particle Separation

Agarose-based SEC media are also commonly used when relatively large biomolecules or biological particles must be separated.

Potential development applications include:

• Large proteins

• Protein complexes

• Polysaccharides

• Macromolecular assemblies

• Virus-related particles

• Other large biological structures

Cytiva positions Sepharose 4 Fast Flow for industrial-scale separation of large molecules and virus particles.

Final suitability for HardGF 3.1 should be confirmed according to its product-specific exclusion characteristics.

Process-Scale Size Exclusion Chromatography

HardGF 3.1 is designed not only for laboratory research but also for scalable chromatography development.

The high-rigidity crosslinked structure can support applications requiring:

• Larger packed columns

• Reproducible bed preparation

• Repeated chromatographic cycles

• Process development

• Pilot-scale purification

• Bulk biomolecule purification

Process-scale agarose SEC media from leading manufacturers similarly emphasize rigid crosslinked matrices for improved flow and large-volume processing.

General Column Packing Workflow

1. Prepare the Resin Slurry

Resuspend HardGF 3.1 thoroughly in a suitable packing or equilibration buffer.

2. Degas if Necessary

Remove excessive trapped air before column packing.

3. Pack the Column

Transfer the homogeneous slurry into the selected chromatography column.

4. Form the Packed Bed

Apply an appropriate packing flow rate according to column dimensions and process requirements.

5. Equilibrate the Resin

Equilibrate the packed bed with several column volumes of the selected mobile phase.

6. Load the Sample

Apply the sample using a controlled loading volume.

7. Perform Size Exclusion Separation

Elute with the selected buffer while maintaining stable flow conditions.

8. Collect Fractions

Collect fractions and analyze the target and impurities using suitable analytical methods.

Factors Affecting SEC Resolution

Important parameters include:

• Particle size

• Column dimensions

• Bed height

• Sample volume

• Sample concentration

• Flow rate

• Buffer composition

• Sample viscosity

• Temperature

• Column packing quality

For size exclusion chromatography, excessive sample volume can reduce resolution.

Method parameters should therefore be optimized according to the target molecule and required throughput.

Sample Loading Considerations

For higher-resolution SEC:

• Use concentrated samples where possible.

• Minimize sample volume.

• Avoid highly viscous samples.

• Clarify samples before loading.

• Maintain consistent column packing.

• Avoid air bubbles.

• Select an appropriate flow rate.

• Monitor pressure during operation.

For preparative applications where throughput is more important than maximum resolution, loading conditions can be adjusted according to process requirements.

HardGF 3.1 vs Conventional Soft Agarose Gel

HardGF 3.1 is designed for users requiring both agarose-based SEC separation and improved bead rigidity.

HardGF 3.1 vs 4% Crosslinked Agarose Media

The agarose percentage alone does not determine chromatographic performance. Crosslinking degree, pore structure, particle-size distribution and manufacturing process also affect selectivity and flow performance.

HardGF 3.1 vs Sepharose 4 Fast Flow Type Media

Cytiva Sepharose 4 Fast Flow is based on approximately 90 μm crosslinked 4% agarose and is positioned for process-scale SEC at relatively high flow rates.

HardGF 3.1 provides:

• 75 μm nominal particle size

• 3.1% agarose

• Multiple-crosslinked structure

• High-rigidity bead design

• Size exclusion chromatography functionality

• SHBC laboratory and bulk supply

HardGF 3.1 should be evaluated using its own separation-range and process-performance data rather than treated as a direct specification equivalent to another commercial resin.

Quality Control for Chromatography Production

Important quality parameters may include:

• Agarose concentration

• Particle-size consistency

• Particle morphology

• Crosslinking consistency

• Mechanical rigidity

• Packed-bed stability

• Flow characteristics

• Pressure-flow behavior

• SEC selectivity

• Batch-to-batch consistency

• Microbial control

• Storage stability

For scale-up applications, chromatographic performance should be evaluated across multiple production batches.

Why Choose SHBC HardGF 3.1?

• 75 μm high-rigidity agarose microspheres

• 3.1% agarose matrix

• Multiple-crosslinked structure

• Designed for size exclusion chromatography

• Suitable for proteins and biomolecules

• Suitable for preparative chromatography

• Suitable for process development

• Laboratory sample evaluation

• Bulk manufacturing capability

• Scale-up supply available

• Custom packaging support

Storage and Handling

Recommended practices include:

• Keep the container tightly sealed.

• Prevent microbial contamination.

• Use clean chromatography equipment.

• Resuspend the medium thoroughly before use.

• Avoid drying hydrated agarose media.

• Avoid aggressive mechanical mixing.

• Use compatible buffers and column materials.

• Validate cleaning and storage conditions for the intended process.

Frequently Asked Questions

What is HardGF 3.1?

HardGF 3.1 is a 75 μm high-rigidity agarose gel filtration medium manufactured from 3.1% agarose through emulsification, washing and multiple crosslinking.

What does HardGF mean?

HardGF refers to SHBC's high-rigidity gel filtration chromatography media series.

What is the particle size?

The nominal particle size of HardGF 3.1 is 75 μm.

What is the agarose concentration?

HardGF 3.1 is manufactured from 3.1% agarose.

What chromatography mode does HardGF 3.1 use?

It is designed for size exclusion chromatography, also known as gel filtration chromatography.

Does HardGF 3.1 bind proteins?

SEC is primarily a non-binding separation method. Separation is based mainly on molecular access to the internal pore structure.

Can HardGF 3.1 be used for antibody purification?

It can be evaluated for size-based antibody purification, aggregate removal and polishing applications.

Can HardGF 3.1 separate protein aggregates?

Yes, aggregate separation is a common application of size exclusion chromatography. Actual resolution depends on the target protein, pore structure, column dimensions and operating conditions.

Can it be used for virus-related purification?

Agarose SEC media are widely used for large biomolecules and virus-related particles. Suitability of HardGF 3.1 should be confirmed using the required target and exclusion characteristics.

What is the molecular-weight fractionation range?

The fractionation range should be based on HardGF 3.1 product-specific test data. It should not be inferred only from the 3.1% agarose concentration.

Is HardGF 3.1 suitable for process scale-up?

Yes. Its high-rigidity, multiple-crosslinked agarose structure is designed for laboratory development and scalable packed-bed chromatography applications.

Can SHBC supply bulk quantities?

Yes. Shanghai SanYu Biotechnology Co., Ltd. supports sample evaluation, pilot-scale requirements and bulk supply of HardGF chromatography media.

Request HardGF 3.1 Samples and Bulk Pricing

Contact Shanghai SanYu Biotechnology Co., Ltd. for HardGF 3.1 samples, technical information and bulk quotations.

Please provide:

• Target protein or biomolecule

• Target molecular size

• Sample type

• Column dimensions

• Required purification scale

• Required resin quantity

• Estimated annual demand

SHBC HardGF 3.1 is designed for research, purification and process-development applications.