Cytiva Butyl Sepharose High Performance Chromatography Resin

When you need reliable polishing and intermediate purification of proteins, the choice of hydrophobic interaction chromatography (HIC) resin directly affects resolution, yield, and scalability. Cytiva Butyl Sepharose High Performance is designed to give you high-resolution separations on a robust, cross-linked agarose matrix that scales smoothly from development to production.

Butyl Sepharose High Performance at a Glance

When you evaluate a HIC resin, you care about resolution, capacity, and how easily it drops into your existing platform process. This resin is built to give you a balanced combination of performance and practicality, so you can improve purity without adding complexity to your workflow.

• Aliphatic butyl ligand on a highly cross-linked agarose base
• High-resolution HIC for intermediate and polishing steps
• Small average bead size (~34 μm) for sharp peaks and good separation
  
• BioProcess-grade resin suitable for scale-up and industrial use

How This HIC Resin Works in Your Purification Workflow

Before you decide where to position Butyl Sepharose High Performance, it helps to understand how the resin interacts with your target molecules. In high-salt conditions, exposed hydrophobic patches on proteins bind to the butyl ligands; as you reduce salt, proteins elute in order of decreasing hydrophobicity.

You can use this mechanism to:

• Separate closely related protein variants or isoforms

• Remove aggregates and hydrophobic impurities after capture or ion exchange

• Fine-tune selectivity by adjusting salt type, concentration, and additives

• Integrate HIC as a polishing step after protein A or other capture steps

By combining HIC with affinity and ion exchange chromatography, you build a multi-dimensional purification strategy that helps you reach your final purity and impurity profiles more consistently.

Key Performance Features You Can Rely On

When you move from screening to production, you need a resin that behaves predictably over many cycles. Butyl Sepharose High Performance is engineered for chemical stability, effective cleaning-in-place, and reproducible column performance, so you can focus on optimizing your process rather than troubleshooting your resin.

This resin offers:

• Cross-linked agarose matrix with particle size 24–44 μm and ~34 μm average bead size for high plate counts and narrow peaks
  
• Working pH range of 3–13 and CIP stability up to pH 14 (NaOH) for robust cleaning and sanitization strategies
  
• Relatively high ligand density to maintain dynamic binding capacity even at lower salt concentrations
  
• Low non-specific binding thanks to the hydrophilic agarose backbone, which supports good recoveries

Together, these features allow you to adjust your HIC step to meet demanding purity and impurity targets while keeping cycle times and buffer usage under control.

Applications: Where This Resin Delivers the Most Value

To get the best return from this resin, you should place it where hydrophobic interactions can help you clean up your product without damaging sensitive biomolecules. It is especially useful when you already have an established capture step and need extra resolving power later in the process.

Typical use cases include:

• Monoclonal antibodies (mAbs): polishing after protein A and ion exchange to reduce aggregates, fragments, and process-related impurities
• Enzymes and other proteins: intermediate purification to separate isoforms, misfolded species, or hydrophobic contaminants
• Peptides and smaller biomolecules: fine resolution of species that differ slightly in hydrophobicity

You can also combine Butyl Sepharose High Performance with other HIC resins (such as phenyl or octyl ligands) to build orthogonal separation strategies when your target is particularly challenging.

Practical Use: Buffers, Loading and Elution Conditions

Once you select this resin, practical questions quickly follow: what buffer conditions to use, how to pack and run the column, and how to balance resolution with cycle time. By standardizing a few parameters, you can quickly move from initial trials to robust, reproducible runs.

In practice, you will typically:

• Equilibrate the column in a high-salt buffer (for example, containing ammonium sulfate or sodium chloride) to promote hydrophobic binding
• Adjust your sample to the same salt concentration before loading to ensure consistent binding behaviour
• Elute with a decreasing salt gradient or stepwise reduction in salt, optionally combined with organic modifiers or pH shifts to fine-tune selectivity
• Clean and regenerate using alkaline solutions such as 1 M NaOH, plus chaotropic agents (urea, guanidine HCl) or organic solvents where appropriate
  

By monitoring pressure, peak shape, and recovery over cycles, you can confirm that the resin maintains performance across your intended cleaning protocol and lifetime.