AdvanceBio RP-mAb

Agilent AdvanceBio RP-mAb Columns

Agilent AdvanceBio RP-mAb columns are purpose-engineered for high-resolution reversed-phase analysis of monoclonal antibodies and large biomolecules. Built on a 3.5 µm superficially porous particle with an enlarged 450 Å pore size, these columns deliver the ideal balance of speed, sensitivity, and robustness for intact mAb, subunit, and fragment characterization.

The wide-pore SPP architecture enhances mass transfer for large proteins, enabling sharper peaks, reduced carryover, and improved quantitation—even for complex mAb therapeutics. Multiple bonded-phase chemistries, including C4, SB-C8, and Diphenyl, provide flexible selectivity for intact, reduced, and stressed-sample analyses.

  • Improved accuracy — 3.5 µm SPP with 450 Å pore increases resolution for intact mAbs and large fragments.
  • Selectable chemistries — C4, SB-C8, and Diphenyl phases optimize retention and selectivity for various mAb workflows.
  • Longer column lifetime — 2 µm inlet frit and rugged packed bed reduce fouling and extend usability.
  • Low carryover — Wide-pore design minimizes adsorption of large proteins and glycoforms.

From intact antibody mapping to Fc/Fab and light/heavy chain separations, AdvanceBio RP-mAb columns provide the reproducible resolution required for biopharmaceutical development, QC, and characterization. Ideal for LC/MS workflows and high-throughput analytical laboratories.

For help selecting the best AdvanceBio column for your mAb workflow, contact Chrom Tech.

Agilent AdvanceBio RP-mAb — Phase Descriptions ▸
AdvanceBio RP-mAb C4
Wide-pore (300 Å) • Bio-inert hardware • Optimized for mAbs • USP L26
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AdvanceBio RP-mAb C4 is engineered specifically for intact monoclonal antibodies and large biotherapeutics. The shorter C4 ligand reduces excessive hydrophobic interaction, improving recovery and peak symmetry for intact proteins while maintaining high resolution.

When to Choose C4

  • Intact mAb profiling
  • LC/MS of large biologics
  • When C8 shows excessive retention
  • When protein recovery is critical

Key Specifications

Separation ModeReversed Phase (Bio-RP)
Pore Size300 Å
Particle DesignSuperficially porous
Application FocusIntact mAbs and large proteins
USP DesignationL26
AdvanceBio RP-mAb Diphenyl
Wide-pore (300 Å) • Aromatic selectivity • mAb variant resolution
▸

AdvanceBio RP-mAb Diphenyl introduces aromatic π-π interactions, providing alternative selectivity versus alkyl phases. It is particularly useful for resolving subtle mAb variants, oxidation states, or closely related protein species.

When to Choose Diphenyl

  • Coeluting mAb variants on C4 or C8
  • Need orthogonal selectivity without changing pH
  • Oxidation/deamidation variant analysis

Key Specifications

Separation ModeReversed Phase (Aromatic selectivity)
Pore Size300 Å
Interaction MechanismHydrophobic + π-π interactions
Application FocusVariant resolution and selectivity tuning
AdvanceBio RP-mAb SB-C8
Wide-pore (300 Å) • StableBond chemistry • Robust low-pH performance • USP L7
▸

AdvanceBio RP-mAb SB-C8 offers a balance of retention and robustness. The C8 ligand provides stronger retention than C4 while maintaining excellent recovery for intact mAbs. StableBond chemistry enhances durability under acidic mobile phases.

When to Choose SB-C8

  • General mAb characterization
  • Need stronger retention than C4
  • Low-pH TFA or formic acid workflows

Key Specifications

Separation ModeReversed Phase (Bio-RP)
Pore Size300 Å
ChemistryStableBond (acid-resistant)
USP DesignationL7

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SKU Name LC Column Phase Particle Size Inner Diameter (ID) Length
SKU
799975-944
Name
799975-944 - AdvanceBio RP-mAb Diphenyl 4.6 x 50 mm, 3.5 u
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
799975-904
Name
799975-904 - AdvanceBio RP-mAb C4 4.6 x 50 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
50 mm
SKU
799775-944
Name
799775-944 - AdvanceBio RP-mAb Diphenyl 2.1 x 50 mm, 3.5um
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
50 mm
SKU
799775-904
Name
799775-904 - AdvanceBio RP-mAb C4 2.1 x 50 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
50 mm
SKU
793975-944
Name
793975-944 - AdvanceBio RP-mAb Diphenyl 4.6 x 150 mm, 3.5
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
150 mm
SKU
793975-904
Name
793975-904 - AdvanceBio RP-mAb C4 4.6 x 150 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
150 mm
SKU
797775-944
Name
797775-944 - AdvanceBio RP-mAb Diphenyl 2.1 x 75 mm, 3.5 u
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
75 mm
SKU
797775-904
Name
797775-904 - AdvanceBio RP-mAb C4 2.1 x 75 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
75 mm
SKU
793775-944
Name
793775-944 - AdvanceBio RP-mAb Diphenyl 2.1 x 150 mm, 3.5
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
150 mm
SKU
793775-904
Name
793775-904 - AdvanceBio RP-mAb C4 2.1 x 150 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
2.1 mm
Length
150 mm
SKU
795975-944
Name
795975-944 - AdvanceBio RP-mAb Diphenyl 4.6 x 100 mm, 3.5
LC Column Phase
Diphenyl
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
100 mm
SKU
795975-904
Name
795975-904 - AdvanceBio RP-mAb C4 4.6 x 100 mm, 3.5 um
LC Column Phase
C4
Particle Size
3.5 um
Inner Diameter (ID)
4.6 mm
Length
100 mm
Key Definitions ▸
AdvanceBio RP-mAb
An Agilent reversed-phase column family optimized for monoclonal antibodies (mAbs) and large proteins, offering selectivity choices (C4, Diphenyl, SB-C8) for intact mAb and related variant workflows.
C4 Phase
A shorter alkyl ligand that provides controlled hydrophobic retention for large proteins. C4 is commonly selected for intact mAb separations when recovery and peak shape are priorities and excessive “stickiness” from longer ligands must be avoided.
SB-C8 (StableBond C8)
A C8 ligand built on Agilent’s StableBond (SB) chemistry, designed for robustness in acidic mobile phases commonly used in protein reversed-phase separations. SB-C8 typically provides stronger retention than C4 while maintaining good recovery for intact mAbs and fragments.
Diphenyl Phase
A stationary phase that adds aromatic (π–π) interactions in addition to hydrophobic retention. Diphenyl is often chosen to change elution order or improve resolution of closely related mAb variants when C4 or C8 does not fully separate critical pairs.
Intact mAb Analysis
A workflow focused on measuring whole antibodies (not digested peptides). Intact methods commonly require wide-pore, bio-optimized phases, high recovery, and stable retention to support biotherapeutic characterization and comparability testing.
Variant / Selectivity Tuning
The practice of changing stationary phase chemistry (C4 vs SB-C8 vs Diphenyl), gradient strength, and temperature to resolve subtle differences between closely related antibody species (variants) without changing the overall LC workflow.
Frequently Asked Questions ▸
What makes AdvanceBio RP-mAb columns ideal for monoclonal antibodies?
AdvanceBio RP-mAb columns are engineered for intact mAbs and large proteins, providing high recovery, strong reproducibility, and optimized selectivity for intact and fragment workflows. The bio-optimized design supports accurate variant and glycoform profiling.
Which phases are available in the AdvanceBio RP-mAb family?
The RP-mAb family includes C4, SB-C8, and Diphenyl phases. These options provide varying hydrophobic and aromatic selectivity for intact mAbs, heavy/light chains, Fc/Fab fragments, and closely related variants.
When should I choose C4 versus SB-C8?
Choose C4 for intact mAbs when recovery and peak symmetry are priorities and excessive hydrophobic retention must be minimized. Choose SB-C8 when stronger retention is needed for variant resolution or when operating in robust acidic mobile-phase conditions.
What advantage does the Diphenyl phase provide?
The Diphenyl phase introduces aromatic (π–π) interactions in addition to hydrophobic retention. It is often selected to change elution order or improve resolution of closely related mAb variants when alkyl phases do not fully separate critical pairs.
What applications benefit most from AdvanceBio RP-mAb columns?
These columns are ideal for intact mAb profiling, reduced mAb (heavy/light chain) analysis, biosimilar comparison, glycoform monitoring, and QC workflows requiring reliable, high-resolution separations of large biomolecules.