Following a round of beta testing by customers looking to avoid the microplumbing of traditional HPLC systems, Agilent Technologies is gearing up to launch a new HPLC chip early next year that reduces the number of fittings and connections by half.
Packed with a traditional protein-separation packing material called Zorbax 5 micrometer SB C-18, the HPLC chip is designed for peptide enrichment and separation. An additional two to three chips packed with different types of separation material will be introduced over the next 12 to 18 months, said Christina Maehr, a spokeswoman for Agilent.
“A main advantage of this integrated system is that by reducing extra tubing you reduce the diffusion of your peptide mixture in liquid when you have a lot of extra column tubing,” said Pierre Thibault, a professor of the University of Montreal Institute for Immunology and Cancer Research, who was one of the beta testers of the new chip. “The net result of extra tubing is lower separation efficiency.”
The new HPLC chip will be the first of its kind to be commercially available when it is released in January 2005. It is as thin as a credit card and measures eight centimeters by about 2.5 centimeters. It works in conjunction with an HPLC-chip/mass spectrometer interface machine called the HPLC-chip cube. The benchtop cube automatically loads and ejects chips, makes hydraulic and electrical connections, switches solvent flows and aligns the emitter tip for nano-electrospraying.
The new chip is expected to cost around $300, and the HPLC-chip cube is expected to cost $30,000. In comparison, conventional nano-LC enrichment columns cost around $600 to $800, and non-electrospray sources cost around $20,000.
The chip is designed to have an average lifetime of seven days if used 24 hours per day. The lifetime of the chip should be taken into account when comparing costs for chips versus conventional systems, Maehr said.
“They system we’ve used has proven rugged. We’ve used it for over 100 analyses without significant loss of performance,” said Thibault. “At the moment it has attractive features, including simplicity, ease of operation and the ability to manipulate small amounts of material.”
Thibault said that the new chip demystifies the microplumbing used with regular HPLC columns. There is reduced chance of leakage and the system is easier to maintain, with less fittings leading to a reduced need for recalibrations.
“Any time you have a switch valve or transfer line, it’s additional plumbing, and additional places where you have to worry about leakage or band broadening or band diffusion,” said Thibault. “You have to try to minimize any peak broadening, and any additional volume that could degrade your performance.”
The downside of the technology is that the type of chromatography available is not very broad at the moment. Also, very limited amount of sample can be loaded on the chip, which can be good if researchers have small amounts of material available.
“This system is ideal for any experiment where you have small amounts of sample available — it could be applied to a drug-discovery program or small molecules,” said Thibault. “It’s exactly the same type of application you have for any LCMS system, except it’s miniaturized.”
Thibault said the new technology should be attractive because of its ease of use for anyone starting up in the proteomics or LCMS area, but that the technology still needs to be assessed more thoroughly.
“People will look at [the HPLC chip] with critical eyes and see if it fits their requirements and fulfills their purposes,” said Thibault.
In addition to releasing its HPLC chip system, Agilent is planning on releasing its Automated Lab on a Chip Platform, or ALP, next month. ALP is an automated version of Agilent’s bioanalyzer, which assesses the quantity and quality of sample on a chip.
The bioanalyzer chip works similarly to gel electrophoresis in that an electric current sucks proteins, DNA, or RNA through etched channels. The smallest particles migrate the fastest, and molecules are separated into portions that appear as peaks.
Launched in 1999, the bioanalyzer’s increase in popularity can be seen through the number of times the chip system has been referenced in literature, said Maehr. Currently, the chip is referenced about 60 times per month in scientific citations, according to Maehr, and Agilent recently sold its millionth chip.
The ALP is a benchtop machine that measures about a meter high. It allows researchers to process thousands of chips per day, rather than the limited 12 chips at a time available with the current briefcase-size bioanalyzer.
“It’s made things a lot easier and saved us a ton of time,” said Kristen Pike, a senior research associate at the National Cancer Institute’s Science Applications International Corporation, who is beta-testing the product. “All the projects we do in the lab are high throughput, so 12 samples per run wasn’t cutting it.”
Pike said so far the only problems with the system are minor glitches with the software. For example, sometimes the software does not import excel files fully, so sample names will have to be typed in manually.
The ALP is expected to cost between $120,000 and $160,000, said Maehr. Agilent’s regular briefcase-size bioanalyzer system costs about $16,000 to $20,000.