How to Choose a Chromatography System: FPLC and AKTA Buyer's Guide for Protein Purification Labs

FPLC and AKTA chromatography systems are the standard platform for automated protein purification in research, biotech, and biopharmaceutical laboratories. Selecting the right system requires understanding your purification scale, column chemistry, and software requirements. This guide covers the key decision points for labs sourcing chromatography systems.

What Is FPLC?

Fast protein liquid chromatography (FPLC) is a low-pressure liquid chromatography technique optimized for protein purification. Unlike HPLC, which operates at high pressures with small-particle columns, FPLC uses larger-particle resins at lower pressures to preserve protein activity and structure. AKTA is the brand name for Cytiva's (formerly GE Healthcare) FPLC system line, which has become the industry standard in research and biopharmaceutical labs.

Key Purification Modes

Affinity chromatography using His-tag, GST, or Protein A/G resins is typically the first step in recombinant protein or antibody purification, providing high selectivity and capture efficiency. Ion exchange chromatography separates proteins based on charge and is used for intermediate purification and polishing. Size exclusion chromatography (SEC) separates by molecular size and is used for final polishing and buffer exchange. Most purification workflows combine two or more of these modes in sequence.

System Scale and Flow Rate

Match the system's flow rate range to your column dimensions and process scale. Bench-scale purification of milligram quantities of protein typically uses columns with 1-5 mL bed volumes at flow rates of 1-5 mL/min. Pilot-scale purification of gram quantities requires larger columns and flow rates of 10-100 mL/min or more. Confirm the system's pump flow rate range covers your current and anticipated future scale.

Detector Configuration

UV absorbance at 280 nm is the standard detector for protein monitoring. Conductivity detection is essential for monitoring salt gradients in ion exchange chromatography. pH monitoring is useful for gradient development and buffer transitions. Confirm the detector configuration matches your purification workflow requirements.

Software: UNICORN and Alternatives

Cytiva's UNICORN software is the industry standard for AKTA system control and method development. Confirm the software version and license status when purchasing refurbished AKTA systems, as older UNICORN versions may not be compatible with current operating systems. Bio-Rad's NGC systems use BioLogic software, which offers a more accessible interface for labs new to automated chromatography.

New vs. Refurbished AKTA Systems

Refurbished AKTA systems from Cytiva are widely available and can offer significant cost savings compared to new instruments. Key checks include pump pressure rating and seal condition, UV detector lamp life and absorbance accuracy, valve and fraction collector function, and software license status. Confirm that the system's pressure rating is compatible with your column chemistry before purchasing.

Source Chromatography Systems from mLab Supply

mLab Supply sources new, refurbished, and surplus FPLC and AKTA chromatography systems from Cytiva, Bio-Rad, and Repligen for research, biotech, and biopharmaceutical laboratories. Submit a quote request and we will confirm current availability, detector configuration, and software license status within 1-2 business days.

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Frequently Asked Questions – FPLC and AKTA Chromatography Buyer's Guide

What is the difference between AKTA Pure, AKTA Start, and AKTA Avant?

AKTA Start is an entry-level system for basic affinity and size exclusion purification at low flow rates. AKTA Pure is the standard research platform with full gradient capability and a wide range of column and detector options. AKTA Avant is a higher-flow pilot-scale system for larger purification volumes. Match the system to your purification scale and column chemistry requirements.

What UNICORN software version do I need for my operating system?

UNICORN 7 and later versions support Windows 10. Older UNICORN versions (5.x, 6.x) may not be compatible with current operating systems. When purchasing a refurbished AKTA system, confirm the included UNICORN version and license type before purchasing.

Can I run ion exchange and size exclusion chromatography on the same AKTA system?

Yes. Most AKTA systems support multiple chromatography modes by changing columns and buffers. Confirm the system's pressure rating is compatible with your column chemistry and that the detector configuration includes conductivity monitoring for ion exchange gradient development.

Can mLab Supply source AKTA systems with specific detector and software configurations?

Yes. Submit a quote request specifying your required AKTA model, detector configuration, UNICORN version, and purification scale and we will confirm available options and lead time.