Keeping Plasma Safe

| Case Study

Haematologic Technologies Inc. Implements Artel’s Liquid Handling Quality Assurance Systems to Ensure Consistent Results

When platelets, red blood cells, and white blood cells are removed from the blood, the clear and protein-rich fluid left behind is blood plasma.  The aptly-named plasma proteins are the proteins found in the blood plasma and play a number of important roles and functions in the human body. Plasma proteins help to regulate the body’s osmotic pressure, which keeps the body’s systems working properly. They also transport various compounds needed by the body, in addition to playing a role in immune system function and blood clotting. An imbalance of plasma proteins can lead a patient to experience symptoms ranging from abnormally dilated blood vessels to a weakened immune system.

article_12_1Modern plasma protein research traces its roots back to the scientifically industrious era immediately following WWII. In the 1940s the U.S. Navy recruited the expertise of Harvard Medical School Professor Edwin J. Cohn to identify a transfusible substance that could be stockpiled in preparation for treating casualties of war. Within a few months, Cohn’s group of researchers developed a methodology to isolate plasma proteins, and not long thereafter utilized their process to successfully isolate the protein albumin from human blood.  The resulting albumin preparations exhibited remarkable effects when administered to patients as an antidote to shock.  The research group’s work represented a major advancement in understanding the biological framework for hematology (the study of the physiology of the blood) and yielded a plasma fractionation process that is still widely used today.  All these factors explain the U.S. Navy’s interest in hematology, and the studies which Dr. Cohn and his group at Harvard were pursuing.

A Vital Role in Medical Manufacturing and Research

In addition to being important to bodily function, plasma proteins are also valuable medical products which are used as components in manufacturing and in research. A number of pharmaceutical products are derived from plasma proteins, with the proteins being either extracted from donor plasma or synthesized in the laboratory. Research involving plasma proteins has resulted in drugs used in treating life-threatening diseases and serious medical conditions such as bleeding disorders, immune system deficiencies, clotting factor deficiencies, burns and shock. Further, research focusing on clinical proteomics−utilizing plasma proteins−identifies proteins and aid in the development of assays for routine and reliable measurement that contribute to solutions for various clinical needs. Assays for a number of disease indicators or biomarkers, which are based on the analysis of single proteins have been FDA-approved and successfully implemented into clinical practice. In cancer research, these include tumor-associated antigen assays for diagnostic use, such as carcinoembryonic antigen (CEA) for colon cancer, prostate-specific antigen (PSA) for prostate cancer, or alpha-fetoprotein (AFP) for testicular and liver cancer.

Today, companies like Vermont-based Haematologic Technologies Inc. (HTI) are making high-quality plasma proteins and reagents readily available to the medical and scientific communities. Given the vital role plasma proteins are increasingly playing in medical and scientific research, suppliers like HTI have developed stringent quality assurance guidelines designed to surpass their customers’ requirements.  Given the critical role of pipettes in handling micro volumes, HTI focuses heavily on quality control for its liquid handling devices.

“For our general manufacturing operations, accurate volume delivery helps us ensure that we are delivering products to our customers with consistent quality, quantity, concentration and specific activity,” said Sara Ethier, HTI’s Quality Assurance Manager. “For our contract testing operations, the accuracy of our pipettes enables us to ensure the accuracy of data that is sent back to the customer.”

A 21st Century Hematologic Lab

HTI boasts a product line of over 150 highly purified and well-characterized proteins including zymogens, enzymes, cofactors, and inhibitors, and also works on stability and release testing—often running anti-drug antibody (ADA) ELISA assays for pharmaceutical companies. Realizing the impact its products have on its customers’ research, HTI (which is ISO 9001:2000-certified) tracks quality assurance in its facilities from basic training to regular equipment calibration. As in many labs, pipettes play an integral role in HTI’s day-to-day operations and Ethier’s department ensures they are not taken for granted, placing a strong emphasis on proper pipetting technique, as well as calibration.

“Our customers have come to expect consistent quality from our products,” said Ethier. “Quality is a crucial part of the work we do, enabling us to provide highly purified, functional and consistent products to our customers. Our equipment and skills are significant parts of this process—a calibrated pipette is not fully functional if it’s being mishandled and a miscalibrated pipette is of little use to the world’s best lab technician. We need to be able to stand behind the high quality of our products, behind the skills of our technicians, and behind the working order of our equipment.”

To help ensure the reliability of their lab processes, HTI turned to Artel, the Maine-based pioneers of liquid handling quality assurance.  The company’s PCS® Pipette Calibration System automatically measures and documents the accuracy and precision of handheld pipettes using dual-dye ratiometric photometry. Because measurement results are traceable to the International System of Units (SI) through National Institute of Standards and Technology (NIST) reference standards, the PCS allows for direct comparison and measurement consistency between operators, methods and pipettes regardless of make, model, manufacturer or location.  As well as being a global standard for liquid volume measurement, hence ensuring that a company’s pipettes in its Singapore lab perform the same as their pipettes in Indianapolis, the PCS is also used as a pipetting technique training tool by labs around the world, providing for consistent results, operator-to-operator, in any lab process involving manual pipetting.

“The pipette is a core fundamental piece of equipment in any lab,” said Bjoern Carle, PhD, Senior Applications Scientist. “In addition to helping ensure the reliability of a laboratory’s tests, the PCS can teach proper pipetting technique because it provides immediate feedback to the user.”

Partners in Lab Quality

To underscore the importance of proper pipetting technique, Artel developed a friendly competition that helps even the most experienced laboratory technicians assess their pipetting skills. The Artel Pipetting Challenge uses the PCS as a tool that goes beyond equipment calibration. At the AACC/Clinical Lab Expo, the Pipetting Challenge competition caught the attention of scientists from HTI, who saw the equipment as a cost- and time-effective solution that supported its stringent quality assurance needs.

HTI had installed a PCS in its laboratory and began using it to ensure the quality of its pipettes and the pipetting technique skills of its technical staff.  “Overall, the use of the PCS helps us ensure the accuracy and reliability of the products that we send to our customers, whether it be a coagulation factor from our manufacturing division or data from our contract testing division,” explained Ethier.

The company seeks to proactively meet regulations ranging from Good Manufacturing Practices (GMP), Good Laboratory Practices (GLP) and ISO 9001. Constant operator training has played an integral role to this approach. Ethier goes on to remark, “We have always trained our laboratory technicians on the proper use of pipettes; it’s a generalized training that covers everything from what solutions you can and can’t use with pipettes, to when to use reverse pipetting. The PCS became an integral part of our training since it allows us to offer our technicians more in-depth training. Once they’ve gone through our general training they can use the PCS to test the skills they learned and identify problem areas. The instrument is always available for employees to practice on.”

And practice they do. During Artel’s 2011 Lab Week Pipetting Challenge, HTI’s lab technicians competed against teams nationwide and won the coveted first place award by scoring the lowest coefficient of variation, showcasing the strength of combining calibrated equipment with efficient pipetting skills.

Putting Training to Use

While proper training is a key element of quality assurance in the lab, it is most effective when paired with strict quality control guidelines. With more than 100 pipettes, HTI uses the PCS on a monthly basis to ensure the instruments are working properly. Using a weekly rotating schedule ensures that all HTI’s technicians have checked the performance of their pipettes at least once each month.

The traditional way to measure pipette volume is to follow the gravimetric approach, which involves weighing water on a balance and converting weight to volume. “When properly controlled, gravimetry is fine for large volume measurement, but when making measurements in the small volume ranges its accuracy and precision fall off drastically,” said Dr. Carle.  “Especially when working below 200 microliters, gravimetric methods often are plagued by evaporation and other environmental issues, which can affect the accuracy of the calibration. And, because measurements often are tracked manually, there is additional room for human error.”

“If you’re working with only a few single-channel pipettes and you’re not dealing in small volumes it’s not too time consuming to calibrate gravimetrically,” noted Ethier. “But when you have a significant number of single-channels pipettes from different manufacturers which are handling small volumes and you start pulling in a few multi-channels, you start looking at significant time consumption.”  Ethier estimates that by integrating the PCS into HTI’s pipette calibration process, technicians have been able to shave off 5-10 minutes per pipette. That amounts to 8-16 hours saved per month—or 96-192 hours per year—which translates into valuable time and labor savings for HTI.

“Before investing in the PCS, we used to send the pipettes back to the manufacturers every six months for calibration. If we thought something was really wrong with an instrument before it was due to be shipped off, we would resort to the balance,” added Ethier, noting the extra time and expense associated with shipping. “It’s an added step that adds up when you’re looking at multiple calibration events per year.”

HTI’s results with the PCS translated into enough benefits for the company to invest in a second Artel system—the MVS® Multichannel Verification System. Like the PCS, the MVS uses dual-dye ratiometric photometry to assess the precision and accuracy of each tip of almost any multichannel pipetting device at volumes as low as 0.01 µL. Often used as a calibration tool for automated liquid handlers, the MVS is employed by HTI to calibrate handheld multi-channel pipettes. “Before running any kind of critical ELISA assays, you want to make sure your pipettes are dispensing properly,” said Ethier. “We weren’t able to do this in the past, but now that we have the ability to do so in-house at a moment’s notice, we can more efficiently deliver on our promises of consistent high quality for our products.”

Life sciences companies stake their reputations on the reliability of their results and the quality of their products. For HTI, which provides the critical tools for cutting-edge medical and scientific research, the burden of quality is appropriately constant given the legacy it upholds. Plasma protein research began as a matter of national security and has evolved into a vital component of life-saving research. Through it all, it was the commitment to high quality standards from researchers companies like HTI and Artel that helped advance the legacy of Dr. Cohn’s discoveries.


About HTI

Haemotologic Technologies, Inc. (HTI) is a biotechnology company specializing in the isolation and characterization of high quality, plasma proteins for in vitro research use, and in the development, validation and implementation of test methods for the pharmaceutical industry.  The company emphasis is focused on proteins involved in the coagulation cascade, as well as the regulation of bone metabolism.  They manufacture and sell over 150 highly purified and well-characterized proteins to over 500 clients worldwide and provide contract services to numerous pharmaceutical companies.  The company serves a worldwide market using a network of international distributors.    For more information about Haematologic Technologies, Inc. or its contract service division, please visit their websites and www.haemtech.com andwww.haemtechbiopharma.com.