Development and production of biopharmaceuticals is a growing segment of pharmaceutical industry. Therapeutic proteins include blood factors, thrombolytic agents, growth factors, interferons, interleukins, therapeutic enzymes and last but not least monoclonal antibodies. More than twenty monoclonal antibody (MAb) therapeutics have been approved for the treatment of a broad range of diseases including autoimmune, cardiovascular, infectious diseases and cancer. A thorough characterization of therapeutic biomolecules is a key task for the successful submission of research and production data for regulatory approvals of new drugs. Due to its high degree of automation, robustness and reproducibility liquid chromatography is the workhorse of the pharmaceutical laboratory. Therefore, HPLC is also frequently used when establishing analysis methods for biotherapeutics. This article features some of the most frequently used HPLC applications for the characterization of biopharmaceuticals.

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This article features some of the most frequently used HPLC applications for the characterization of biopharmaceuticals.

Characterizing Biotherapeutics with HPLC — Smart Solutions for Large Molecules

A round-up of the winners at this year’s HTC Awards.

A round-up of the winners at this year's HTC Awards.

The HTC Awards

Modern chromatographic analysis is costly, in both time and resource requirements, and it is subject to many internal and external pressures to limit the cost of analysis, identify and quantify impurities at lower levels and increase sample throughput.

Laboratories today, especially those performing more routine analysis, have a stated desire to increase productivity (sample throughput) and decrease cost per analysis. One proposed solution has been to move to smaller HPLC particles (sub-2 µm) and faster flow-rates. This solution, while effective, produces high operating pressures beyond the capability of most of the conventionally installed HPLC instruments, requiring an investment in new HPLC instruments with high pressure pumping capabilities (up to 1200 bar) to realize the benefits these fully porous sub-2 µm columns can provide. These newer instruments, generally referred to as UHPLC systems, offer some real benefits in terms of reduced system volume and have been designed to achieve optimum performance from the growing number of fully porous sub-2 µm columns being introduced by column manufacturers. However, the cost of replacing the large number of conventional HPLC instruments installed in laboratories to realize the benefits these sub-2 µm columns can provide would be high and results in questions about the cost-effectiveness of such a strategy.

Recently, a newly developed Kinetex 2.6 μm core-shell particle has been commercialized that offers the performance benefits of fully porous sub-2 μm particles but at substantially lower operating pressures. Columns packed with 2.6 μm core-shell particles can provide outstanding efficiency on conventional HPLC instruments. Because 2.6 μm core-shell columns do not require high pressure pumping systems, conventional HPLC instruments with operating pressure limits less than 600 bar can be used to realize productivity increases and analysis cost reductions. 

We demonstrate here that Kinetex core-shell columns can elevate the performance of conventional HPLC systems to a level that is comparable with UHPLC systems. This can be easily achieved by the implementation of straightforward adjustments to conventional HPLC systems that reduce system volume.1 When combined with increased detector response times these optimized HPLC systems can accommodate the much narrower peak widths obtained with core-shell particles. The benefits of the system optimization process are illustrated for the determination of impurities and related substances for acetylsalicylic acid (European Pharmacopoeia Monograph 0309).

Demonstrating core-shell columns that can elevate the performance of conventional HPLC systems to a level that is comparable with UHPLC systems.

Increased Productivity and UHPLC Performance from Conventional HPLC Systems using Core-shell Particle Columns

Validation is not an easy task. It is an ongoing complex process that evaluates the entire, sample specific process, products and analytical methods and consists of four qualification phases: Design Qualification (DQ), Installation Qualification (IQ), Operational Qualification/Performance Verification (OQ/PV) and Performance Qualification (PQ).

Before the first sample is analysed, users have to verify and document that the system meets the user requirement specifications, is correctly installed and working according to the manufacturer’s specifications in the selected environment.

Validation of the system is complete after the instrument hardware, software, data, analytical method and operators are factored in the process. In addition to simple performance tests for each isolated component, it is essential to confirm that the complete system works according to the performance specifications of the analysis method (holistic approach). Here it is important to take the specific requirements of the applied method into account. This ensures that all critical parameters influencing the results are identified and properly monitored over the system’s life cycle.

Validation is not an easy task. It is an ongoing complex process that evaluates the entire, sample specific process, products and analytical methods.

The Column-03-05-2010

Characterizing Biotherapeutics with HPLC — Smart Solutions for Large Molecules

The HTC Awards

Increased Productivity and UHPLC Performance from Conventional HPLC Systems using Core-shell Particle Columns

How to Validate a GPC/SEC System