Bio SEC-5

Agilent Bio SEC-5

Agilent Bio SEC-5 HPLC columns are engineered for high-resolution size exclusion chromatography of large biomolecules, complex mixtures, and high–molecular weight species. With tightly controlled pore structures and a proprietary neutral hydrophilic surface, Bio SEC-5 columns minimize secondary interactions to deliver exceptional accuracy across a wide range of sample types.

Designed for rugged, reproducible performance, Bio SEC-5 columns provide outstanding stability under high-pH, high-salt, and low-salt conditions—making them ideal for method development, QC workflows, and advanced biopharmaceutical characterization. The enhanced pore volume increases peak capacity and resolution, enabling clear separation of aggregates, fragments, and intact biomolecules with confidence.

  • Exceptional resolution for large proteins, vaccines, and high-MW biomolecules
  • Proprietary hydrophilic surface minimizes nonspecific interactions
  • Rugged silica construction delivers excellent column lifetime and batch-to-batch reproducibility
  • Stable under high-pH and high-salt mobile phases for flexible method optimization
  • Pore sizes up to 2000 Ã… support analysis of very large complexes

Chrom Tech offers the full Bio SEC-5 portfolio in multiple pore sizes and formats to support vaccines, monoclonal antibody characterization, and other high-molecular-weight applications. For help selecting the optimal pore size or column dimension for your workflow, our technical support team is here to assist.

Agilent Bio SEC-5 — Phase Description ▸
Bio SEC-5
Size Exclusion Chromatography • 5 µm Silica Particles • Diol-Bonded • Multiple Pore Sizes
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Agilent Bio SEC-5 columns are robust size exclusion chromatography (SEC) columns packed with 5 µm silica particles and a hydrophilic diol-bonded surface. They are engineered for reliable, reproducible separation of proteins, monoclonal antibodies (mAbs), aggregates, and fragments under aqueous conditions.

When to Choose Bio SEC-5
  • You require rugged, routine SEC analysis
  • You need lower backpressure on standard HPLC systems
  • You analyze proteins, mAbs, aggregates, or biosimilars
  • You prioritize robustness and column lifetime over maximum efficiency
How It Compares

Compared to Bio SEC-3 (3 µm), Bio SEC-5 (5 µm) offers reduced system backpressure and increased durability, making it ideal for routine QC workflows. While resolution may be slightly lower than 3 µm formats, it provides dependable aggregate quantitation and sizing performance.

Key Specifications
Separation Mode Size Exclusion Chromatography (SEC)
Particle Size 5 µm
Surface Chemistry Hydrophilic diol-bonded silica
Pore Sizes Available Optimized for peptides, proteins, and monoclonal antibodies (varies by molecular weight range)
Mobile Phase Aqueous buffers (phosphate, saline, or compatible systems)
Primary Applications Aggregate analysis, fragment profiling, biosimilar comparison, protein purity testing

Practical note: Maintain appropriate buffer ionic strength to suppress ionic interactions and ensure accurate size-based separation.

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SKU Name LC Column Phase Particle Size Inner Diameter (ID) Length
SKU
5190-6866
Name
5190-6866 - Agilent SEC-5,500A,21.2x300mm
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
21.2 mm
Length
300 mm
SKU
5190-6865
Name
5190-6865 - Agilent SEC-5,300A,21.2x300mm
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
21.2 mm
Length
300 mm
SKU
5190-6864
Name
5190-6864 - Agilent SEC-5,150A,21.2x300mm
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
21.2 mm
Length
300 mm
SKU
5190-6863
Name
5190-6863 - Agilent SEC-5,100A,21.2x300mm
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
21.2 mm
Length
300 mm
SKU
5190-6862
Name
5190-6862 - Agient Bio SEC-5 2000A, 4.6x50mm guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-6861
Name
5190-6861 - Agilent Bio SEC-5 1000A, 4.6x50mm guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-6860
Name
5190-6860 - Agilent Bio SEC-5 500A, 4.6x50mm guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-6859
Name
5190-6859 - Agilent Bio SEC-5 300A, 4.6x50mm, guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-6858
Name
5190-6858 - Agilent Bio SEC-5 150A, 4.6x50mm, guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-6857
Name
5190-6857 - Agilent Bio SEC-5 100A, 4.6x50mm, guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
5190-2545
Name
5190-2545 - Bio SEC-5, 2000A 7.8x50mm, 5um Guard
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
7.8 mm
Length
50 mm
SKU
5190-2544
Name
5190-2544 - Agilent SEC-5, 2000A, 4.6x150mm
LC Column Phase
SEC
Particle Size
5 um
Inner Diameter (ID)
4.6 mm
Length
150 mm
Key Definitions â–¸
Size Exclusion Chromatography (SEC)
A chromatographic technique that separates biomolecules based on hydrodynamic size. Larger molecules elute first, while smaller molecules penetrate pore volume and elute later.
5 µm Particle Size
A larger particle diameter that reduces system backpressure and enhances robustness. Compared to 3 µm formats, 5 µm particles provide durable performance for routine QC workflows.
Diol-Bonded Silica
A hydrophilic surface modification applied to silica particles that minimizes secondary hydrophobic or ionic interactions, improving protein recovery and maintaining native structure.
Protein Aggregates
High-molecular-weight species formed through protein association. SEC is commonly used to quantify aggregates during stability, comparability, and regulatory testing.
Hydrodynamic Radius
The effective molecular size of a compound in solution. SEC separations are governed by hydrodynamic radius rather than polarity or hydrophobicity.
Frequently Asked Questions â–¸
What is Bio SEC-5 primarily used for?
Bio SEC-5 columns are used for size-based separation of proteins, monoclonal antibodies (mAbs), aggregates, and fragments. They are commonly applied in QC testing, stability studies, and biosimilar comparison workflows.
How does Bio SEC-5 compare to Bio SEC-3?
Bio SEC-5 uses 5 µm particles, resulting in lower backpressure and enhanced robustness compared to 3 µm formats. Bio SEC-3 provides higher resolution, while Bio SEC-5 is ideal for routine analysis and longer column lifetime.
Why is buffer composition important in SEC analysis?
Proper ionic strength and pH control suppress unwanted electrostatic interactions between proteins and the stationary phase. This ensures separation is governed strictly by molecular size.
What pore size should I select?
Larger pore sizes are recommended for high-molecular-weight proteins and mAbs, while smaller pore sizes are more suitable for peptides and smaller proteins. Matching pore size to analyte size improves resolution.
Is Bio SEC-5 suitable for regulated environments?
Yes. Bio SEC-5 columns are widely used in regulated QC laboratories for aggregate quantitation, purity assessment, and lot-to-lot comparability due to their robustness and reproducible performance.