Single-use technologies are established as a go-to for making genomic medicines, including the viral vectors used for gene delivery.
But how do you purify small volumes for preclinical and clinical studies without losing too much to processing or leaving capacity unused?
Download this app note to discover a method that has:
- Small hold-up volume and flow rate support making it efficient at the purification of small drug volumes
- Single-use flow path with an option for aseptic connectors to maintain a closed flow path
- A small benchtop footprint making efficient use of valuable facility space
ÄKTA ready™ 450 system is a single-use liquid chromatography system designed for small-scale manufacturing (Fig 1). To meet the needs of purifying small drug volumes, the system has a low flow range of 5 to 450 mL/min per pump and a low hold-up volume. The single-use flow path minimizes the need for cleaning and cleaning validation, allowing for quick changeover between productions and reducing the risk of carryover. The ÄKTA ready 450 chromatography system offers the performance and documentation required for use in a GMP-regulated environment. Designed for small-scale manufacturing: • Small hold-up volume and flow rate support efficient purification of small drug volumes. • Single-use flow path with option for aseptic connectors to maintain a closed flow path. • Small benchtop footprint makes efficient use of valuable facility space. Designed for small-scale GMP manufacturing We designed the ÄKTA ready 450 chromatography system for purifying small drug volumes efficiently. All wetted parts of the liquid flow path are single use, minimizing the risk of crosscontamination. Flow kit installation is fast and easy to help minimize risk of error, reduce time spent on non-value-adding activities, and support short turnaround times between batches. Many products are sensitive to contamination from the surrounding environment. In other cases, the drugs themselves can be very potent or even toxic, so avoiding exposure to the environment is also important. A closed process facilitates preparation and increases safety in both instances. The system’s small footprint enables efficient use of facility cleanroom space. The system has a benchtop design, and all valves, monitors and tubings are mounted on the forward-facing wet side of the system for easy access. The space in front can hold columns of volumes up to 2.5 L. It can also accommodate a column holder for smaller columns. The system is designed for cleanroom environments and to withstand harsh sanitization. The ÄKTA ready 450 chromatography system is controlled by external human-machine interface (HMI) with UNICORN™ control software, which adheres to GAMP™ 5 practices and is 21 CFR Part 11 compliant. Intuitive method editing enables quick recipe creation. The system has a standard for open platform communication (OPC), and the interface is compatible with thirdparty system control software. Fig 1. The ÄKTA ready 450 chromatography system is single-use and has a low flow rate and hold-up volume for efficient purification of small quantities in a GMP environment. 1 2 4 3 3 5 7 6 8 9 10 1 6 2 4 5 3 Inlets Inlet valve block Flow meters Pump A Pump B Pre-filter position Main valve block with air trap and sensors pH and UV sensors Outlet valve block Outlets 7 8 10 9 2 Flow kit ÄKTA ready 450 chromatography systems can be used for isocratic or gradient elution. Flow kits are available for both types of operation, either with Tri-Clamp™ or AseptiQuik™ connectors. Markers show where to connect the sensors, and the valve blocks snap into place for easy attachment to the system. Sensors do not require calibration. An installation test confirms that the system has been set up correctly. Installation can be done in as few as 10 minutes. Flow kits (Fig 2) are produced in an ISO class 7 clean room, double-bagged, and gamma-irradiated before delivery. Upon completion of the run or batch, the single-use flow path is disposed to prevent carryover between production batches or campaigns, facilitating processing in a multiproduct facility. Flexibility and manufacturing capability The system flow path has twelve inlets (eight are on pump A and four are on pump B for gradient elution) and six outlets. Flow is measured for each pump. The flow path also features an automated air trap and a location for a pre-filter, and each can be individually bypassed. The air trap automatically detects and prevents introduction of air into the column. An inline air sensor protects the chromatography column from air. The pressure drop over the pre-filter and column can be measured individually. Post-column sensors monitor pressure, conductivity, temperature, pH, and absorbance (UV sensor). The UV sensor can measure up to three wavelengths between 206 nm and 700 nm simultaneously. The absorbance (A) ratio between two wavelengths (e.g., A260 /A280 ) can be measured and used for pooling criteria. Fig 2. Flow kit for ÄKTA ready 450 chromatography system. The flow kit includes pump head(s), valve blocks, and sensors. Fig 3. ÄKTA ready 450 flow chart. AS Inlets Outlets Pu C P AS L L P P C pH UV W W
