Cytiva AKTA avant 25 Protein Purification System

Overview

The Cytiva AKTA avant 25 is a fully automated, benchtop preparative chromatography system engineered for robust method development, process optimization, and scalable purification of recombinant proteins, monoclonal antibodies (mAbs), viral vectors, and other biopharmaceutical molecules. Based on the proven principles of high-performance liquid chromatography (HPLC) and low-pressure liquid chromatography (LPLC), the AKTA avant 25 delivers precise gradient formation, real-time multi-parameter monitoring (UV, conductivity, pH, temperature, pressure), and intelligent fraction collection—all under unified control via UNICORN 6 software. Its modular architecture supports affinity, ion exchange, size exclusion, hydrophobic interaction, and multimodal chromatography modes. Designed specifically for early-stage process development in R&D labs and pilot-scale transfer, the system bridges the gap between analytical screening and GMP-compliant manufacturing—enabling direct scalability from HiScreen columns to production-scale columns without re-optimization.

Key Features

• BufferPro™ Online Buffer Preparation: Integrated four-channel valve system enables automatic, real-time mixing of stock solutions (acid/base, salt, water) to generate precise buffer compositions across wide pH and ionic strength ranges—eliminating manual titration, reducing preparation time by ~30 minutes per condition, and improving reproducibility and scalability.
• Design of Experiments (DoE) Integration: Embedded DoE module in UNICORN 6 allows simultaneous variation of multiple critical parameters (e.g., gradient slope, flow rate, load density, wash stringency), significantly reducing experimental runs while maximizing information yield for statistical modeling and robustness assessment.
• Intelligent Column Protection: Dual inline pressure sensors (pre- and post-column) continuously monitor ΔP across the packed bed; dynamic pressure limits prevent overloading of Capto™, MabSelect™, or other BioProcess™ resins—ensuring column integrity and long-term performance stability.
• UNICORN 6 Software Platform: Compliant with FDA 21 CFR Part 11, featuring electronic signatures, role-based access control, immutable audit trails, and secure database storage. Includes Method Editor for intuitive drag-and-drop protocol building, Column Logbook for full traceability, and UniTag™ QR-code scanning for automatic column history registration.
• Flexible Fraction Collection: Built-in fraction collector supports 3–96-well formats with programmable volume thresholds, temperature-controlled racks (4–25 °C), auto-detection of rack type, and real-time tube position verification—minimizing sample loss and cross-contamination risk.
• Enhanced System Diagnostics: Integrated bubble sensors at sample inlet and pump heads, plus self-calibrating UV and conductivity detectors, ensure continuous operational integrity and early fault detection during unattended runs.

Sample Compatibility & Compliance

The AKTA avant 25 is validated for use with all major Cytiva prepacked columns—including HiScreen™, HiTrap™, Tricorn™, and XK™ series—as well as user-packed columns up to 26 mm internal diameter. It supports standard biotherapeutic feedstocks such as clarified mammalian cell culture harvests, bacterial lysates, and plasma-derived fractions. All hardware and software components meet ISO 9001 quality management standards and are designed to support GLP/GMP environments. Full regulatory documentation—including IQ/OQ/PQ protocols, 21 CFR Part 11 validation packages, and vendor-supplied test reports—is available through Cytiva’s Regulatory Support Services. The system complies with IEC 61010-1 for laboratory equipment safety and EN 61326-1 for electromagnetic compatibility.

Software & Data Management

UNICORN 6 serves as the central operating and data governance platform. It provides centralized, encrypted database storage for all methods, raw sensor data, chromatograms, fraction metadata, and audit logs—retained in immutable format with timestamped versioning. Each user account is assigned granular permissions (e.g., method edit, run execution, data export, system configuration), enforced via password authentication and optional two-factor login. Electronic signatures are required for critical actions including method activation, batch release, and audit trail review. All changes trigger automatic entries in the system audit trail, recording operator ID, timestamp, action type, and prior/post values. Data export adheres to ALCOA+ principles (Attributable, Legible, Contemporaneous, Original, Accurate, Complete, Consistent, Enduring, Available) and supports CSV, PDF, and native .u6d formats compatible with LIMS and ELN integration.

Applications

• Monoclonal antibody purification workflows (Protein A capture → low-pH viral inactivation → cation exchange polishing)
• Process parameter screening and optimization for bispecifics, Fc-fusion proteins, and antibody-drug conjugates (ADCs)
• Development of scalable purification strategies compliant with ICH Q5A(R2) and Q5B guidelines
• Characterization of resin lifetime, binding capacity, and cleaning-in-place (CIP) efficiency
• High-throughput purification of CRISPR-Cas9 complexes, lentiviral vectors, and extracellular vesicles
• Supporting Quality-by-Design (QbD) initiatives through DoE-driven design space definition and control strategy implementation

FAQ

What is the maximum recommended flow rate for the AKTA avant 25 system?

The system is rated for continuous operation up to 25 mL/min, with pressure capability up to 20 MPa (200 bar), suitable for high-resolution separations on small- to medium-bore columns.
Can UNICORN 6 be used to control multiple AKTA systems simultaneously?

Yes—UNICORN 6 supports networked deployment across up to four connected instruments via Ethernet, enabling synchronized method execution and centralized data aggregation.
Does the AKTA avant 25 support pH gradient elution?

Yes—via BufferPro™ integration, the system enables precise, real-time pH modulation using acid/base stock solutions, with automatic compensation for temperature- and concentration-dependent pKa shifts.
Is column tracking mandatory when using UNICORN 6?

Column tracking via UniTag™ QR codes or manual entry is optional but strongly recommended for full traceability, especially in regulated environments where column usage history impacts process validation and change control.
How does the system handle buffer depletion during extended runs?

Integrated level sensors in solvent reservoirs trigger configurable alerts and can automatically pause runs or switch to backup channels if primary buffers fall below user-defined thresholds.
What types of detectors are included as standard?

The base configuration includes a dual-wavelength UV/Vis detector (280 nm and 254 nm), a conductivity detector, a pH sensor, and a temperature probe—all calibrated and verified per manufacturer specifications before shipment.