Bio SEC-3

Agilent Bio SEC-3 Columns

Agilent Bio SEC-3 columns are designed for high-resolution size exclusion chromatography of proteins, peptides, and biomolecular complexes—especially when pairing SEC with mass spectrometry (SEC-MS). Built on 3 Âµm, high-efficiency silica particles, Bio SEC-3 columns deliver sharp, symmetrical peaks and exceptional separation of monomers, aggregates, and fragments.

Engineered for modern bioanalysis, Bio SEC-3 columns are fully compatible with denaturing organic/aqueous mobile phases commonly used in SEC-MS workflows, providing reliable performance whether operating with low-salt MS-friendly conditions or higher-salt SEC buffers. Their mechanical robustness and optimized pore structure enable accurate, reproducible molecular-weight determinations across a wide range of biotherapeutic modalities—including mAbs, ADCs, peptides, and other next-generation biologics.

With full scalability from analytical to preparative dimensions, Bio SEC-3 columns support method development, routine QC, and high-throughput characterization on any LC/MS platform. For labs requiring sensitivity, reproducibility, and dependable molecular-size profiling, Agilent Bio SEC-3 delivers industry-leading performance and confidence.

Agilent Bio SEC-3 — Phase Description ▸
Bio SEC-3
Size Exclusion Chromatography • 3 µm Silica Particles • Diol-Bonded • Multiple Pore Sizes
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Agilent Bio SEC-3 columns are high-efficiency size exclusion chromatography (SEC) columns packed with 3 µm silica particles and a hydrophilic diol surface. They are engineered for high-resolution separation of proteins, peptides, monoclonal antibodies, and aggregates under aqueous conditions.

When to Choose Bio SEC-3
  • You need high-resolution aggregate analysis
  • You require improved efficiency with 3 µm particles
  • You analyze proteins, mAbs, fragments, or biosimilars
  • You need reproducible QC performance with minimal adsorption
How It Compares

Compared to polymer-based SEC columns, Bio SEC-3 provides the efficiency advantages of small-particle silica. The diol-bonded surface minimizes secondary interactions while maintaining robust performance in standard aqueous buffer systems.

Key Specifications
Separation Mode Size Exclusion Chromatography (SEC)
Particle Size 3 µm
Surface Chemistry Hydrophilic diol-bonded silica
Pore Sizes Available Optimized for proteins, peptides, and monoclonal antibodies (varies by molecular weight range)
Mobile Phase Aqueous buffers (phosphate, saline, or compatible systems)
Primary Applications Aggregate quantitation, fragment analysis, biosimilar comparison, protein purity assessment

Practical note: Maintaining proper ionic strength in the mobile phase is essential to suppress ionic interactions and ensure true size-based separation.

Product List

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SKU Name LC Column Phase Particle Size Inner Diameter (ID) Length
SKU
5190-2501
Name
5190-2501 - Bio SEC-3, 100A 7.8x300mm, 3um Column
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
300 mm
SKU
5190-2515
Name
5190-2515 - Bio SEC-3, 300A 7.8x50mm, 3um Guard
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
50 mm
SKU
5190-2514
Name
5190-2514 - Agilent SEC-3, 300A, 4.6x150mm
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
4.6 mm
Length
150 mm
SKU
5190-2513
Name
5190-2513 - Bio SEC-3, 300A 4.6x300mm, 3um Column
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
4.6 mm
Length
300 mm
SKU
5190-2512
Name
5190-2512 - Agilent SEC-3, 300A, 7.8x150mm
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
150 mm
SKU
5190-2511
Name
5190-2511 - Bio SEC-3, 300A 7.8x300mm, 3um Column
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
300 mm
SKU
5190-2510
Name
5190-2510 - Bio SEC-3, 150A 7.8x50mm, 3um Guard
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
50 mm
SKU
5190-2509
Name
5190-2509 - Agilent SEC-3, 150A, 4.6x150mm
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
4.6 mm
Length
150 mm
SKU
5190-2508
Name
5190-2508 - Bio SEC-3, 150A 4.6x300mm, 3um Column
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
4.6 mm
Length
300 mm
SKU
5190-2507
Name
5190-2507 - Agilent SEC-3, 150A, 7.8x150mm
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
150 mm
SKU
5190-2506
Name
5190-2506 - Bio SEC-3, 150A 7.8x300mm, 3um Column
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
300 mm
SKU
5190-2505
Name
5190-2505 - Bio SEC-3, 100A 7.8x50mm, 3um Guard
LC Column Phase
SEC
Particle Size
3 um
Inner Diameter (ID)
7.8 mm
Length
50 mm
Key Definitions â–¸
Size Exclusion Chromatography (SEC)
A chromatographic technique that separates biomolecules based on hydrodynamic size rather than chemical interaction. Larger molecules elute first, while smaller molecules access pore volume and elute later.
3 µm Particle Size
Smaller particle diameter that increases column efficiency and resolution. In SEC, 3 µm particles improve aggregate and fragment separation compared to larger-particle packings.
Diol-Bonded Silica
A hydrophilic surface treatment applied to silica particles that minimizes secondary hydrophobic or ionic interactions, helping maintain native protein structure and improving recovery.
Protein Aggregates
High-molecular-weight species formed by association of protein molecules. SEC methods are commonly used to quantify aggregates during biopharmaceutical development and quality control testing.
Hydrodynamic Radius
The effective size a molecule presents in solution, determined by molecular weight and three-dimensional structure. SEC separations are governed by hydrodynamic radius rather than polarity.
Frequently Asked Questions â–¸
What is Bio SEC-3 primarily used for?
Bio SEC-3 columns are designed for size-based separation of proteins, monoclonal antibodies (mAbs), aggregates, and fragments. They are widely used for aggregate quantitation and purity analysis in biopharmaceutical development and QC workflows.
How does Bio SEC-3 differ from polymer-based SEC columns?
Bio SEC-3 uses 3 µm silica particles with a diol-bonded surface, offering high efficiency and resolution. Polymer-based columns provide broader pH stability, while Bio SEC-3 emphasizes efficiency and reproducibility in standard aqueous buffer systems.
Why is ionic strength important in SEC?
Adequate buffer ionic strength suppresses unwanted ionic interactions between proteins and the stationary phase. This ensures separation is governed strictly by molecular size rather than electrostatic effects.
What pore size should I choose for my protein?
Larger pore sizes are recommended for high-molecular-weight proteins and mAbs, while smaller pore sizes are better suited for peptides and smaller proteins. Matching pore size to molecular weight improves resolution.
Is Bio SEC-3 suitable for regulatory and QC environments?
Yes. Bio SEC-3 columns are widely used in quality control, stability testing, and biosimilar comparison studies, offering reproducible performance and high-resolution aggregate analysis.