HPLC-Chip Team Creates Industry-First Platform
Presenters and winners: Neil Cook, Turtle Brennen, Hongfeng Yin,
Karen Seaward, Kevin Killeen, Darlene Solomon
“We thank Hongfeng, Kevin, Turtle and Karen for their willingness to take big technical risks that had the potential for significant business outcomes; for creating an entirely new Agilent platform; and for contributing great technology that has helped our LC-MS platforms become a market leading Agilent business!” said Darlene Solomon, Agilent chief technology officer.
"I am extremely proud and pleased that the HPLC-Chip team won the Barney Oliver Prize," said Neil Cook, vice president and director, Agilent Laboratories. "They demonstrated key factors required to create a novel platform for Agilent: vision, determination, stamina, innovation, and effective collaborations with internal and external partners. The result is a clearly differentiated product and technology platform that will continue to provide tremendous customer value in future years. Congratulations to everyone who shared and supported the vision of bringing this microfluidics platform to the market."
In the following interview Hongfeng, Kevin, Turtle and Karen offer their perspectives on winning the 2009 Barney Oliver Prize for Innovation.
What is the HPLC-Chip/MS?
Kevin: The HPLC-Chip/MS is a microfluidic device -- about the size of a credit card – that simplifies the study of proteins and other biochemical samples. It makes very complex technology accessible to a wider group of users. The HPLC-Chip/MS combines two processes: 1) high performance liquid chromatography (HPLC) that separates compounds so they can be identified and quantified, and 2) nanospray ionization for mass spectrometry (MS) that determines the composition of a molecule. Using the HPLC-Chip, scientists can identify samples within minutes to a couple of hours with high sensitivity and reproducibility. Previous methods suffered from low sample throughput and were difficult to reproduce.
Why do customers like the HPLC-Chip/MS?
Kevin: The chip integrates everything from sample preparation to nanospray, so it saves time and provides a high degree of precision and confidence in the measurement. In a traditional work flow, all the connections between components and internal surfaces can potentially cause sample loss; each step has some degree of uncertainty even in the hands of very well qualified researchers, so leaks and errors can reduce the precision of the measurement. The chip integrates these components and works consistently over and over again. It really excels when sample volumes are limited, like drug profiles extracted from a sample of a few hairs or examining a minimal amount of bone from a 5,000 year old mummy.
What’s your reaction to winning the Barney Oliver Prize?
Karen: There’s no question that getting the Barney Oliver Prize is a real thrill and honor because it’s very highly regarded and competitive. You never think your work is going to make it to that level, not because of the work itself, but because a lot of things have to come together to meet the criteria for this prize. We don’t have control over the many factors that are involved. For example, I started working on the HPLC-Chip in 2000; Agilent introduced it as a product in 2005, and we received the award in 2009. There is a lot of time between these steps. In addition, the innovation has to make a significant amount of money for the company. The chip project was pushed along with collaborations, publications and by getting top people in the field involved, but you never really know what will make it successful. A lot of advances have been made along the way in the LCMS product line, in the chip customization, in the robustness of the manufacturing and in the user acceptance. The HPLC-Chip has found its place.
Turtle: The Barney Oliver Award was given to four people, but these four people could not have succeeded on their own. End of story. We relied heavily on a lot of other people from all over the place – throughout Labs, in Agilent sites in Waldbronn, Germany; and Little Falls, Delaware; and in outside companies that we worked with at various points. We hired people to do original work with us who provided technologies we didn’t have in house. We couldn’t have done it on our own.
What motivated you to work on this project?
Hongfeng: We have had lots of fun working on the HPLC-Chip technology. We have a great team with expertise in many areas, and we respect each other. I feel that I can contribute to the project and learn a lot from other team members at the same time. It’s rewarding to see that we can make something happen, something that none of our competitors can do, even five years after our products became commercially available.
Karen: I got involved on a dare. One of the big issues was the chip lamination process. We needed to have the layers of the chip stick together well enough to put significant pressure (3000psi) into the chromatography column. A team member didn’t think we’d be able to reach that target. So the dare was on making the lamination process work.
Of course, I didn’t know if the lamination would work either, but I had extensive background in gas-phase plasma processing from earlier work that I did on compound semiconductors. I knew that this technology had been used to increase adhesion between a wide variety of materials. Some key observations by team members led us to the plasma parameters that needed to be optimized. It turned out that observing, and later understanding the nuances made a big difference.
Turtle: I’ve loved working on this project precisely because it’s a long-term project with a compelling vision. What motivates me are the day to day discoveries, questions and solutions that come up.
How would you describe your team?
Karen: The group that developed this chip from 2000 to 2003 was a collection of half a dozen Ph.D.s working on their own special areas. Folks took the parts they knew how to do, got those parts working well, and then we put them together. There was a high level of focus: Hongfeng on the chip packing and applications, Turtle on the valve interface, Kevin on the laser ablation of the structure, as well as large contributions by other team members at the time.
Here is an example. One team member did video rate imaging of the spray coming out of the tip, looked simultaneously at electrical parameters, and figured out what conditions would produce a stable spray. One day he wanted to show me what he had found. His lab was a totally different world from the one I worked in. With his experimental setup, you could watch the spray and learn why it was becoming unstable. His work and discussions with others led to our development of a plasma-deposited coating to make the electro spray tip work. I don’t think anyone anticipated the importance of the tip coating until after the imaging had been done.
Turtle: A great thing about working here is that everybody is smart, curious and curious. Take both meanings of the word “curious,” or eccentric or opinionated or however you want to put it. We’re an eclectic bunch, so we have fascinating people, fascinating projects. We question things and solve them. We fail sometimes, but usually we don’t call it failure. We just go, “Oh, that didn’t work. Let’s go over here.”
I question what makes a Labs employee. What makes people working in Labs any different? When you talk to people, you just know: that’s somebody who is an Agilent Labs type person. I think curiosity is what differentiates. You look and go, “Hmmm, I wonder why that’s like that?” People don’t do that every day. It’s just this subtle thing. “Why does that work? How does that work?” It’s not a question of “Oh, I wonder how I can solve that.” It’s more a question of “What’s going on?” I’m in the middle of that environment, and I’m a cat in the cream.
Kevin: Management support is also really essential. When it looks like we’ve got great technology, something disruptive, these are very fragile phases of a project. I can’t say enough about how all the managers really made this move forward and allowed it to happen with constant dialogue and collaboration.
What other people worked with you?
Turtle: Our transfer team -- Agilent people in Waldbronn who commercialized the HPLC-Chip technology -- supported the work for years before transfer and kept their finger on the pulse the whole time. There was a champion for the project, and they contributed engineering time. Their involvement was invaluable.
Kevin: Waldbronn started with a very aggressive timeline and thought they could pull off the transfer of our technology in nine months. They made it! That doesn’t just happen because you have good technology. It happens because of the people involved, and like all good success stories in Labs and throughout Agilent, it really is the people who make it happen. All successful transfers from Labs to an Agilent business take regular communications and follow up visits. Our teams in California and Waldbronn visited back and forth, and what we learned over there, we’d bring back to Labs and vice versa. What we learn from these visits continues to drive our innovation and success and still allows us to move forward on new chip designs today.
Hongfeng: Over the course of technology transfer and product development, we have had many internal partners and external collaborators. In addition to our transfer team in Waldbronn, we work with our LC/MS group in Santa Clara to develop the MS interface. Agilent’s columns and supplies group in Little Falls gave us lots advice on packing material, and today we are also working very closely with the Stratagene team on another packing material.
We also are very fortunate to have many excellent external collaborators. They validated our technology, gave us advice on new applications and helped us introduce our technology to other thought leaders in the field.
Did Agilent have a competitive advantage in developing the HPLC-Chip/MS?
Hongfeng: Agilent has a unique competitive advantage in that we have strong core technology in laser ablation and micromaching, and domain knowledge in chromatography and mass spectrometry applications. There have been debates on whether one needs to focus on technology or on applications. If you focus on just the technology, say a hammer, then everything looks like a nail to you. On the other hand, it is not a good situation either when you have a unique application problem without the technology expertise to solve it.
How does this project compare with others at Agilent Research Laboratories?
Turtle: This is a perfect example of a Labs project because the final application went in a direction we weren’t expecting - towards LC/MS. It’s one of those classic, far-reaching projects where you think you know what you’re going to end up with, but you don’t. The result in the end is probably more valuable, has a bigger market potential and goes further with the original perceived advantages than you expected. This has been the absolute archetypal Labs project.
What about your results?
Kevin: We ended up with something that we’re really proud of today. It’s still seen as a benchmark in the industry, so much so that we’ve inspired copycats. Our key competitors are coming after us, and their imitation is the highest form of flattery. Also, Agilent’s emerging and growing portfolio of mass spectrometers really allows us to develop all these new applications.
Turtle: We were working on something that other people were working on too. We came out with our product first, and the quality of our microfluidics still beats the heck out of the microfluidics that other people are doing in the same field. Our work is still cutting edge, right on the front of what can be done with LC/MS, microfluidics, low flow rates.
Karen: I have a curious Barney Oliver story to share. In my second year at HP Labs, I met Barney when he made the rounds at the annual science fair. At the time we were developing plasma processing for compound semiconductors, and I was explaining to Barney some plasma work that I had done. He asked if we made the plasma tool that I was showing. I said that we did not, and he left my poster after that one question. So here we are 25 years later, and I’m getting the Barney Oliver award in his honor for plasma work. Thank you, Barney! We have completed the circle.
“In the analytical instrument business it is rare to have a unique product that is so dramatically different from convention and with such a strong value proposition. The business results have been incredible as the HPLC Chip has been an integral part of helping make the LCMS business one of the fastest growing businesses at Agilent.” Patrick Carberry, Agilent Senior Director Mass Spectrometry Programs, Agilent Life Sciences Group
“The HPLC-Chip technology made it possible to establish collaborations for the first time with key thought leaders in the area of proteomics and glycomics, and it has significantly changed the way many of these collaboration partners and other customers are analyzing biological samples today.” Rudi Grimm, Director of Science and Technology, Agilent Life Sciences Group
"The chip changed the course of my research. It's not biology that leads progress, but technology that allows us to explore biology and make progress." Professor Carlito Lebrilla, Department of Chemistry, University of California, Davis
“The HPLC-Chip/MS is a very, very potent competitive differentiator. Agilent has engineered the complexity out of the product and created the unprecedented ability to do plug-n-play nanospray LC/MS. Superb robustness and precision allow comparative and multi-lab collaborative studies and have opened up large potential markets such as biomarker discovery and validation, and clinical protein assays. When customers see this absolutely unique level of performance, precision and ease of use, they’re really blown away.” Ken Miller, Global Senior Marketing Manager, LC/MS, Agilent Life Sciences Group
“The HPLC-Chip team had the vision more than 15 years ago that this concept would build Agilent’s future. Now they are expediting the market penetration of the HPLC-Chip-MS system by developing new applications, establishing fast prototyping and test capabilities in their lab, and delivering HPLC chip prototypes to leading researchers in life science who mandate top-notch bio-analytical measurement tools. Fred Strohmeier, VP and GM, Liquid Phase Analysis Division, Agilent Life Sciences Group
“The HPLC-chip/MS solution is an excellent example of how new technology can enable more researchers to do what was considered for experts only. Disruptive technologies often begin within interdisciplinary research groups like Agilent Labs, where the foundation for the HPLC-chip was laid. Working closely with the product development teams, to turn the vision into a truly differentiated solution, was the key to the incredibly positive feedback we have received from our customers.” Tom van de Goor, R&D Section Manager, Liquid Phase Separations Business, Agilent Life Sciences Group
We are living through an unprecedented revolution in the life sciences that is driven by the development and application of new measurement methods. One battlefront in this revolution is the application of microfabrication techniques to chemical and biological measurements. The HPLC-Chip/MS team has capitalized on a diverse background to establish Agilent as the leading provider of microfluidics-based separation techniques for biochemical analysis by mass spectrometry. The resulting solution enables enhanced performance and ease of use while conserving the customer’s precious experimental samples.
This technology has transformed nano-electrospray ionization from a laborious technique done by only a few highly trained scientists to a routine analysis capability that can be performed by any mass spectroscopist. For several years, Agilent has been the only company able to offer these capabilities, which has helped to fuel our enormous recent success in mass spectrometer sales.
Beginning of microfluidics research
Microfluidics research at HP Laboratories and Agilent Research Laboratories dates back to 1992, roughly coinciding with the beginning of such investigations at universities and national laboratories. In the early phases, essentially all external research was centered on chemical separation by capillary electrophoresis. This has been the dominant approach for researchers because it requires only the application of high voltage across the channels to direct flow and to achieve chemical separation using the differential electrical mobility of analytes in a liquid buffer. When Agilent was founded in 1999, the only commercially available microfluidic device for chemical analysis was the glass-based Caliper/Agilent Bioanalyzer product. The Bioanalyzer performs capillary electrophoresis in a gel matrix to separate and quantitate labeled RNA, DNA, and proteins via optical fluorescence.
The Agilent Labs team took a distinctively different approach to the problem. They focused on pressure-driven flows, using laser ablation to create polymer microfluidic devices to perform chemical separation and to introduce the analytes for detection by electrospray mass spectrometry (LC/MS). High-performance liquid chromatography (HPLC) is the most widely used method for analyzing chemical mixtures. A solution of chemicals is forced through a column containing a solid material with differential affinity for the analyte molecules. The solid phase is densely packed, so high pressure is necessary to drive the liquid solution through the column.
First requirement for success
The first requirement for success of the HPLC-Chip/MS technology was to create a microfluidic device that can withstand the large pressures employed in HPLC (up to 400 atmospheres). A major challenge was to create a leak-free connection to the device that allowed flow switching, and to achieve this with minimal dead volume. To realize the promise of microfluidics, the sample size and flow rates must be very small. With miniscule flow rates, even a tiny amount of dead volume can destroy the performance.
The team’s creative insight was to sandwich the microfluidic device between the faces of a rotary valve. The valve “stator” interface solves the problem of applying pressure via capillary connections, and the “rotor” interface allows a reliable face seal for fluid switching with multiple and simultaneous connections between engraved loops in the rotor and ports to channels in the microfluidic device. Thechip-to-valve interface solved numerous technical problems, all within the small confines of the microfluidic chip and valve. Without this elegant solution, researchers in other laboratories struggled with these requirements, often employing bulky connectors with large dead volumes.
Power of multidisciplinary research
The success of this work demonstrates the power of multidisciplinary research. The original project was a collaboration between the life science lab, bringing expertise in chemistry and chemical analysis, and the electronics lab, bringing expertise in materials and microfabrication. Once Hongfeng and Kevin conceived the basic idea, they faced some major challenges. First, a method had to be found to form the flow channels and shape the electrospray tip. They chose UV laser ablation as the fabrication technique. For the base material, they chose polyimide, which is inert and robust to the solvents and pressures used in HPLC.
A unique, high precision CAD-CAM laser ablation system was built and optimized to pattern individual layers and to create the chip electrospray tip. Next, a method was needed for the reproducible, high-adhesion, lamination of polyimide sheets to form a structure tolerant of high pressure. This was addressed with O2 plasma “activation” of the patterned polyimide films in a Labs-built vacuum laminator. Next came the development of a reproducible and automated way to pack the column, followed by the creation of a zero-dead volume metalized fluid contact junction for biasing a non-etting plasma coated electrospray tip. The final piece of the invention was the crucial chip interface to the mass spectrometer that created a precise positioning and easy to use, high pressure, low dead volume rotary valve interface to the HPLC-Chip-/MS device.
The 2009 Barney Oliver Prize for Innovation is for the conception and execution of an integrated polymeric microfluidic HPLC-Chip-/MS device, the design of its method for fabrication and the demonstration of its superiority in performance over conventional nano-LC columns plus discrete spray tips; and for successfully transferring the technology to Agilent’s bio-analytical measurement business to enable creation of the HPLC-Chip/MS product. The HPLC-Chip/MS technology has created unique collaboration opportunities for Labs and business engagement for Agilent.
In summary, the team pioneered the application of microfluidics to systems combining chromatographic separation with mass spectrometry. They applied their diverse backgrounds to solve many challenges with innovative approaches. They integrated sample enrichment, chemical separation, and electrospray ionization to achieve enhanced performance over conventional techniques, with dramatic improvements in ease of use, reproducibility, and quality. In the process they helped to establish Agilent as the leader in microfluidics separation technologies, and contributed to the outstanding growth of the company’s high end mass spectrometry sales. Their work is a testament to the culture of innovation and business contribution that Barney Oliver established as the first director of Hewlett-Packard Laboratories, and that continues at Agilent Research Laboratories.
About the Barney Oliver Prize for Innovation
The Barney Oliver Prize for Innovation honors Bernard M. Oliver (1916 – 1995), who was a scientist, inventor, and innovator. Agilent Research Laboratories have awarded the prize annually since 1999 for contributions to Agilent that result from work done in the Labs and that demonstrate Barney's outstanding qualities of creativity, innovation, technical depth, breadth of expertise, and respect for business value. The prize consists of $10,000 after-tax cash and a bronze statue.
Dr. Oliver, known to all as Barney, was a man of enormous intellect, curiosity and vision. When he was 19, he graduated from Stanford University with a B.A. in electrical engineering. A year later he completed an M.S. from the California Institute of Technology, where he earned a Ph.D., graduating magna cum laude at the age of 24.
Barney then joined Bell Telephone Laboratories where he quickly established a reputation for brilliant, creative insights and clever inventions. In 1952, Bill Hewlett and David Packard persuaded Barney, whom they had known since their student days, to join their fledgling operation as director of research. In 1957 he became vice president of Research and Development, and in 1966, he established Hewlett-Packard Laboratories, which he directed until his retirement in 1981.
During his career Barney was awarded 60 patents and authored 71 papers that reflect a remarkable breadth and depth of thought, ideas, and actions spanning physics, mathematics, electronic and electrical engineering, education, and social issues. He was active in the IEEE and served as its president in 1965.
Barney had a lifelong interest in astronomy, and in particular the use of radio telescopes for the search for extraterrestrial intelligence. Between 1982 and 1993 he was the chief engineer of the SETI (Search for Extraterrestrial Intelligence) Institute and member of the SETI board of trustees.
Barney received many awards. In 1986 President Reagan awarded Barney the National Medal of Science for “translating the most profound discoveries of physical and communication science into the electronic, radio, and computer systems which have improved our culture and enriched the lives of all Americans.” In 1997 the SETI Institute established the Bernard M. Oliver Chair, and in 2004 Barney was inducted posthumously into the National Inventors Hall of Fame. Among his academic honors were the Halley Lectureship on Astronomy and Terrestrial Magnetism of Oxford University (1984).