Cytiva Mono S 5/50 GL Strong Cation Exchange Column

Mono S 5/50 GL is Cytiva’s high-resolution strong cation exchange column designed for fast, reliable protein separations in small-scale process development, polishing steps, and method optimization. With a 1 mL bed volume and robust MonoBeads resin, it helps you resolve closely related charge variants with confidence.

Product Overview: What Is Mono S 5/50 GL?

When you need precise control over protein charge-based separations, Mono S 5/50 GL gives you a compact, high-performance option. It is a prepacked strong cation exchange column, ideal for analytical runs and small preparative purifications on systems such as ÄKTA chromatography platforms.

Mono S 5/50 GL is packed with monodisperse, rigid polystyrene/divinyl benzene beads functionalized with sulfonate groups (S), providing strong cation exchange behavior over a broad pH range. The column has inner dimensions of 5 × 50 mm and a 1 mL bed volume, enabling sharp peaks at short run times when you work within a recommended flow range up to 3 mL/min and column pressure up to 4 MPa (40 bar).

Key Features and Benefits for Your Protein Purification

To decide whether Mono S 5/50 GL is the right IEX column for your workflow, it helps to see how its technical properties translate into practical advantages. The points below summarize how this strong cation exchanger supports robust, reproducible separations in research and bioprocess development.

• High resolution in a small format – The 10 µm monodisperse bead size and optimized column geometry give you narrow peak shapes and excellent separation of proteins with subtle charge differences, such as isoforms or post-translational variants.
• Fast method screening and polishing – Short bed height and 1 mL volume allow rapid gradients, so you can screen buffers and conditions quickly or perform polishing steps without long run times.
  
• Robust performance and long lifetime – The rigid matrix tolerates high flow rates and cleaning in place with up to 2 M NaOH and a wide range of organic solvents, supporting many cycles when you follow recommended cleaning protocols.
  
• Wide chemical and pH compatibility – Mono S 5/50 GL can be operated in commonly used aqueous buffers from pH 2–12 and in the presence of chaotropes such as urea or guanidine hydrochloride, giving you flexibility to handle challenging samples.
  
• Easy integration with ÄKTA systems – The column is delivered with 1/16'' connectors suitable for standard FPLC tubing, making it simple to connect, test, and transfer methods across labs and systems.

Typical Applications and Use Cases

Once you know the basic specs, the next question is how this column behaves with real samples. Mono S 5/50 GL is widely used for protein polishing, charge variant analysis, and method development where you need high resolution in a compact format.

You can use the column to purify positively charged proteins at the working pH, including enzymes, recombinant proteins, and monoclonal antibodies that require charge-based polishing. It is also suitable for separating hemoglobin variants such as HbA1c, where the method demands precise resolution of closely eluting peaks.

Typical use cases include:

• Final polishing of recombinant proteins after affinity or capture steps
• Separation and profiling of mAb charge variants in development workflows
• Optimization of buffer conditions and gradients before scaling to larger strong cation exchangers
• Analytical quality control runs to confirm batch-to-batch consistency

How Mono S 5/50 GL Fits into Your IEX Workflow

In most ion exchange workflows, you are not using Mono S 5/50 GL in isolation; instead, it sits alongside affinity, other IEX steps, and size exclusion chromatography. Understanding its place in the sequence helps you design a robust purification strategy.

A common sequence would start with capture on Protein A or His-tag affinity media, followed by a polishing step on a strong cation exchanger such as Mono S to resolve closely related species. You can then finish with gel filtration or desalting to remove small molecules and adjust the final buffer. On an ÄKTA system, this type of workflow is straightforward to automate with preprogrammed gradients and column scouting.

Product Range and Ordering Information

When you standardize methods across projects or want a clear handover to colleagues, it is useful to have the exact product codes at hand. The table below summarizes Mono S 5/50 GL and related Mono S columns so that you can plan capacity and scaling options from the outset.

Product name	Type	Column dimensions (ID × L)	Bed volume	Cytiva code / SKU
Mono S 5/50 GL	Strong cation exchanger	5 × 50 mm	1 mL	17-5168-01
Mono S 10/100 GL	Strong cation exchanger	10 × 100 mm	8 mL	17-5169-01
Mono S 4.6/100 PE	Strong cation exchanger	4.6 × 100 mm	1.7 mL	17-5180-01

Selection Guide: When to Choose Mono S 5/50 GL

Choosing between Mono S and other Cytiva IEX columns usually comes down to your protein’s charge, sample volume, and required throughput. A concise decision framework can save you time when you design or optimize a purification strategy.

Use the guidelines below as a quick reference:

Choose Mono S (strong cation exchanger) when:

• Your target protein is positively charged at the chosen pH (typically at least one pH unit below its pI).
• You need high-resolution separation of charge variants or closely related isoforms.
• You want fast gradients and short run times at small scale.

Choose Mono Q (strong anion exchanger) when:

• Your protein is negatively charged at the working pH (≥ 1 pH unit above pI).
• Early scouting suggests better selectivity on anion exchange media.

Choose larger Mono S or RESOURCE/Capto columns when:

• You have already optimized conditions on Mono S 5/50 GL.
• You need to process larger sample volumes while keeping comparable selectivity and performance.
  

This simple selection logic helps you position Mono S 5/50 GL as a scouting and polishing tool, while pointing the way to scale-up columns with similar chemistry.