Optimists of every stripe are hoping that new science and technology applications will once again emerge to rescue us from the jaws of the current economic grinder. The million dollar questions are: Which technologies and what applications?
Nanotechnology is often eyed as the white knight, and even greater hope is heaped on nanobiotechnology.
The Pew Charitable Trusts, one of the top supporters of investigations in this field, views nanotech as "the practice of manipulating and manufacturing things on the atomic scale."
At leading research institutions globally, there is considerable interest in the applicability of nanotechnology across numerous disciplines. According to Pew literature, nanotechnology "has the potential to change everything about our daily lives—cars, clothing, food or more efficient solar cells."
To be sure, there is some concern with technology on this tiny scale. Along with the Woodrow Wilson International Center for Scholars, Pew initiated the Project on Emerging Nanotechnologies in 2005 to help minimize potential health and environmental risks and to ensure public involvement while helping to realize the benefits of nanotechnology.
Still, hope about this technology abounds. When 700 of the world's sharpest minds gathered at the World Economic Forum in Dubai late last year, Jackie Ying, executive director of the Singapore-based Institute of Bioengineering and Nanotechnology, pointed to areas where the impact may be greatest: early detection of communicable diseases to stave off pandemics, increased production of pure drinking water, and new forms of energy.
Focusing on the biopharmaceutical area, Patricia Young, senior marketing product manager at Waters Corp., says, "Biopharmaceutical researchers are not generally sample-limited. They can usually access large amounts of samples from their bioreactors. They may be monitoring the media to ensure sufficient nutrients remain, waste products do not build up, or the amount and integrity of the biological product they're producing."
When samples are abundant, the preferred analytical approach is usually analytical-scale ultra performance liquid chromatography (UPLC) on sub-2-micron particles with columns featuring larger internal diameters, typically 2.1 mm. However, for early discovery or biomarker analyses, researchers are frequently working with small amounts of sample, often less than a picomole. In these cases, the best approach is nanoscale UPLC on 75-micron ID columns to minimize sample losses and increase sensitivity.
Waters currently offers the ACQUITY UPLC for analytical scale separations, which was introduced in 2004. Following that, the company launched its nanoACQUITY UPLC system, which uses small columns with internal diameters of 75-microns to 1 mm.
"The rationale for the nanoscale system," says Young, "is that it allows the injection of a small amount of analytes without suffering sensitivity losses through band broadening.
"Following the nanoACQUITY UPLC, we added the TRIZAIC UPLC module. This scalable microfluidic device removes the need for user intervention in creating the fittings for nanoscale chromatography," she says.
On the production side, analysis for the most part is done with optical detection on tunable UV or photodiode array detectors. "When there's a problem, however, and there is something totally different in the expected profile, such as an unexpected protein signature or a new peak in a peptide map, there is a need to revert to the power tools, the mass spectrometers, to figure out the precise events in the bioreactor," says Young.
"The nanoACQUITY UPLC system has been shipping since 2004, and we have TRIZAIC for increased usability," she adds. "We will be shipping in the summer of 2009.
"We took a bit more time with this because we reevaluated the marketplace and realized that the nanoTile substrate we were using was great for 75-micron ID, but we needed to expand it for use with 300-micron ID and above to address a wider range of molecules.
"They both sound very small, but the application spaces are markedly different. The 75-micron-type nanoTile for TRIZAIC is for strict proteomics applications. If you scale up to 300-micron ID, then the applicable areas include metabolism and DMPK samples, which are important to biotech and pharmaceutical researchers," Young explains.
"That's where we are now—we're gaining an understanding about where people want to use TRIZAIC at nanoscale and capillary flow rates." Young adds that, "TRIZAIC is the future of Waters' separations strategy. We're starting in nanoscale and capillary, and then expanding into analytical flow rates. That's our goal."
Providing some sense of the delivery times on these systems, Young says that the TRIZAIC UPLC system will be available this year. "We need to prove that there's a marked increase in ease of use to compell people to go from our very robust chemistries in the packed tube to the TRIZAIC. We're experiencing that transition now in the capillary nano-flow area."
"That will be the challenge—building systems with consistency that will run continuously," explains Adams. He sees consistency of manufacture as one of the key challenges going forward with potential nanoscale instruments.
Zeroing in on an available nano HPLC system that Shimadzu currently offers—the Prominence, Adams says that "customers doing proteomics analyses have a limited amount of sample, and they typically want to get the maximum sensitivity out of that sample.
"We know that by scaling down an HPLC system, they can get more sensitivity due to the sensitivity of the columns, with sharper peaks, but we also know that when using mass spectrometry and nanospray, we can get the entire spray to go into the mass spec—allowing us to sample the whole sample instead of a portion of it," says Adams. "Proteomics researchers who are trying to increase sensitivity are the ones who will use this kind of system."
While developing the Prominence HPLC system, which is the platform for everything within the HPLC line, "we had an engineer at the National Institutes of Health (NIH) who was charged with taking our capillary system and optimizing it. While he was doing that, he also tried to produce a nanosystem using our off-the-shelf hardware while also evaluating all of the other available nanosystems," explains Adams.
"His conclusion was that we can't make a good nanosystem—one that was reproducible from existing equipment. We needed to do something better.
"When he went back to Japan, those systems opened up and led to the nano system we offer today—the Prominence Nano LC. So, we gained first-hand experience from the user perspective, from the exposure our engineer received at the NIH who was then supported by a great engineering team at Shimadzu Corp. in Japan," says Adams.
He adds that the end result has been the creation of a robust, easy-to-use nanosystem with reproducible results that is also easy to maintain and can run in a 2-D configuration—all at analytical flow rates and nano-flow rates.
This flexibility is good for customers who may want to use nano one day. "When they figure out what they really need, with this system they'll be able to switch over and run a capillary or microflow rates," he says.
Shimadzu launched two new nano systems at Pittcon 2009—a new proprietary particle size measurement method and single nano particle analyzer. The company also announced the launch of a new BioSpec-nano UV-Vis spectrophotometer for life science applications.
Dayin Lin, product manager, HPLC-Chip/MS, Agilent Technologies, says there is considerable interest in nanotechnology, which allows the use of very small samples—a necessity in some cases. A strong following has already been built around miniaturized technologies, such as the Agilent 2100 series bioanalyzer, which has become a fixture in DNA analysis and a dominant tool for RNA analysis in the course of the past 10 years.
On the LC side, Agilent was the first company to commercialize the micro fluidic device—the Agilent 1200 Series HPLC-Chip/MS. Now there are a number of applications for proteomics—LC-MS using nano-techniques, says Lin.
"We're expanding beyond protein identification and into DMPK (drug metabolism and pharmacokinetic) studies," he adds. Agilent has an ultrahigh capacity version of the 1200 Series HPLC Chip, which is being tested in collaboration with Novartis. It is being employed in analyses where the amount of sample is extremely small, derived from serial bleeding, and sometimes only a drop of mouse blood is available. The HPLC-chip/MS requires only 0.1 to 4 µg of a sample for a proteomic or metabolomic analysis.
According to Lin, the specific technology involved in the nano-flow LC-MS uses nanospray. "What we have in the case of this particular product, called the HPLC chip, is the only available fully integrated micro fluidic LC-MS device.
"Integrated into this device are micro connections, columns, as well as other fittings, all of which are packaged together in a credit-card-sized chip device into which samples can be injected. An LC is used to drive the flow and, at the end, an electrospray ionization flows out of this HPLC chip. This spray is highly compatible and may be analyzed using any system from the entire Agilent LC-MS line, that is, whether they are single- or triple-quadrupole systems," says Lin. This simplifies several systems that are challenging to deal with separately.
"This is what researchers are after," concludes Lin. "This approach simplifies the method and connects all the systems while saving time, streamlining the workflow process and resulting in more successful experiments."
• For more information, contact:
Brian Murphy, PR, Waters Corp., 508-482-2614, firstname.lastname@example.org
Kevin McLaughlin, PR, Shimadzu Scientific Instruments, 800-477-1227, email@example.com
Stuart Matlow, PR, Agilent Technologies, 408-553-7191, firstname.lastname@example.org