January 15, 2007 (Vol. 27, No. 2)

Miniaturization of Fluidics and Accuracy in Biorepositories Top Product Offerings

One of the highlights of the upcoming “LabAutomation2007” conference in Palm Springs, CA, will be an emphasis on emerging technologies. While numerous sessions will focus on a variety of approaches, the key areas of concern appear to be adjusting technology to meet smaller and smaller size requirements, finding ways to avoid contamination in expensive or irreplaceable samples, and bringing pharmaceutical research closer to the realm of clinical practice.

Spurred on by the issuance of pharmacogenomics guidance by the FDA and the introduction of the Genomics and Personalized Medicine Act of 2006, molecular diagnostics is becoming an integral part of disease management and therapy in patient stratification, drug regimen selection, toxicity avoidance, therapeutic monitoring, and detection of disease predisposition, according to Shuqi Chen, Ph.D., CEO of IQuum (www.iquum.com).

Testing often needs to be conducted in near-patient settings to provide “actionable diagnostic information,” says Lingjun Chen, associate director of operations at IQuum. The company’s Liat™ (Lab in a Tube) analyzer enables nucleic acid testing at the bedside or a clinical trial site by minimally trained personnel in less than an hour.

“Liat takes nucleic acid testing out of the environmentally controlled lab, puts it into the general lab or at the bedside, and allows anyone to perform these sophisticated tests,” says Dr. Chen. “By putting integrated amplification and detection in one vessel, we’ve reduced the possibility of contamination and the need for manual processing and training.”

The Liat sample vessel is comprised of a flexible tube that contains all assay reagents prepacked in tube segments separated by peelable seals. Multiple sample processing actuators in the Liat system compress the tube to selectively release reagents from tube segments, move the sample from one segment to another, and control reaction conditions.

IQuum is leveraging this technology to develop tools for biological sample testing. The company is also applying the Liat technology to biodefense, including the development of the Bioagent Autonomous Network Detector in collaboration with the U.S. Department of Homeland Security for bio-aerosol monitoring.

Small But Mighty

“Miniaturization has the potential to revolutionize the field of analytical chemistry,” notes Sammy S. Datwani, Ph.D., senior scientist of advanced technologies at Eksigent Technologies(www.eksigent.com). “In order to miniaturize HPLC, one must address the need for microfabrication methods, flow control methods, connections, injectors, and reliable, low-volume detectors that are compatible with the pressures on the order of hundreds of atmospheres. Eksigent’s technology tightly integrates the critical components of HPLC—a decades-old technology—into a microfabricated format, specifically the injector, separation column, pumps, valves, and detector.”

The goal Dr. Datwani says, “is to realize the promise of microfluidics through exceptionally high reproducibility, customization, automation, reduced system complexity, increased ease of use, increased throughput, and reduced costs for solvents and reagents.” Work on the chip-based system was supported by the National Institute of Standards and Technology’s advanced technology program.

“The technology has the capability to accelerate chemical analysis, specifically for drug development, which is paramount to aiding big pharma in discovering drugs faster and bringing next generation drugs to market sooner,” Dr. Datwani adds. “There is an economic impact from accelerated drug development due to fast separations, high quality, and reliable data. The close coupling and tight integration of system components leads to reduced system dispersion and improved data quality of the overall system. Multiplexing could result in savings in labor, reagents, solvents, and space.”

Gary Valaskovic, Ph.D., founder and president of New Objective (www.newobjective.com) believes that the miniaturization of LC-MS-based analysis holds promise for applications in biomarker determination and general bioanalysis. He cautions that a key technical challenge is the efficient coupling of microfluidic elements.

Conventional LC-MS fittings that have been scaled down for use with capillary tubing make routine use a challenge. Misaligned tubing and significant extra-column volume can cause damaged tubing ends, system clogging, or column failure.

Dr. Valaskovic explains, “The PicoClear™ microfluidic capillary connection system will impact distinct areas of laboratory automation by miniaturization of laboratory components and, ultimately, instruments and assays. We like to think of it as the functional equivalent of solder in interfacing capillary-scale liquid components. Our immediate market for the product is nanobore LC-MS as applied to qualitative proteomics and quantitative biomarker analysis.”

The PicoClear fluoropolymer union-core contains fritted, fused-silica tubing inserted into the inlet end of the union. It is used as an in-line, metal-free filter. The ability to validate the connections allows monitoring of the filter for contaminants. Reliable operation in excess of 9,000 psi has been demonstrated, according to Dr. Valaskovic. The coupling elements were tested for chromatographic performance with columns ranging from 20–75 mm.

A technological alliance between the Paul Scherrer Institute (www.psi.ch) of Switzerland and Vitalea Science (www.vitaleascience.com)is developing a compact accelerator that will change the way drugs are tested in clinical trials, according to Stephen Dueker, Ph.D., president of Vitalea Science. The new instrument, BioMICADAS, is a compact particle accelerator that could remove radioactivity from clinical drug trials.

BioMICADAS measures radioactive drugs at microdoses that would even be safe for studies in pregnant women and infants. While current, early-stage procedures using animal models to test toxicity often have poor correlation with drug action in humans, procedures using the BioMICADAS will enable evaluations in human subjects without overexposure to radioactive drugs, according to the company.

“BioMICADAS is a new version of accelerator mass spectrometry (AMS), a form of isotope-ratio mass spectrometry developed for carbon dating, that now traces carbon fourteen biochemicals and drugs with sensitivity and precision,” notes Dr. Dueker. “This sensitivity enables microdosing, where microgram quantities of a development drug are tested in people under an exploratory IND that requires a minimal animal-toxicology package. It is designed to allow early human evaluations of drug candidates using harmlessly small doses.”

AMS is the engine behind microdosing. Attomole sensitivity with carbon-14-labeled substrates enables mg/Kg human dosing as well as conventional ADME evaluations using 1,000–10,000 times less radioactivity, according to the companies.

Rethinking Repositories

“Biobanks are expected to play an important role both within traditional pharmaceutical research and in long-term community-based studies,” say Simon Sheard, technical sales manager at RTS Life Science (www.rtslifescience.com). “Industrial-quality robotics are critical in biobanking applications where there is often only a single opportunity to collect and process that sample.”

Biomaterials can cover a wide range of sample types. Pharmaceutical companies operating biorepositories may store tumor/specific tissue samples for particular studies, whereas biorepositories, such as the U.K. Biobank and the HUNT Biobank in Norway, collect, process, and store blood samples from members of the general public for future research. While blood can be collected, dried, and stored easily at room temperature, there are limitations on the subsequent uses for the collected samples. One alternative is to extract the DNA from whole blood and store this at -20ºC. However, this is an expensive and time-consuming process.

RTS combines the Assay Platform™ technology developed for screening applications with vision technology to create a fully automated system capable of centrifuging containers of blood, identifying the heights of the various fractions, and generating liquid-handling protocols to accurately aspirate the various fractions and dispense them into suitable formats for future storage.

This technology, currently going through the final stages of testing at customer sites, is capable of accurate processing at 10–20 times the speed of a laboratory scientist and offers all the usual advantages of automation, including improved sample tracking, ability to process at low temperature, improved safety, and full and automatic data integration with the client’s LIMS system, Sheard says.

A second RTS biobanking application at Bristol University in the U.K. gives researchers a ready supply of genomic DNA. On receipt of a sample, it is “immortalized” and stored. When needed, a small collection of cells is removed from storage and “grown-up,” Sheard says.

“Collecting many different genotypes will help to explain biological pathways and speed up the identification of new substances that might lead to personalized medicine,” says Dietmar Reisch, Ph.D., product manager at REMP (www.remp.com).

“But those new methods are extremely sensitive to differences in sample quality. Recently, it has been shown that proteins in plasma that undergo multiple freeze/thaw cycles exhibit severe damage as quickly as the second cycle. However, plasma proteins stored at –70°C for up to four years showed no significant protein degradation. Limiting freeze/thaw cycles therefore seems more important to maintaining the integrity of the plasma proteome than degradation caused by long-term storage.”

Pfizer (www.pfizer.com) is taking advantage of an automated, –80°C sample-store, Dr. Reisch says. Samples can easily be traced and will be collected from robots 24/7. Researchers can identify samples of patients that belong to a group of interest within seconds by searching the database with predefined filters and can arrange those samples for ready use in an experiment without reformatting steps. Experiments can be done on demand. Sample integrity is improved, which allows researchers to compare datasets that might be separated by years. The ability to reuse expensive samples will help to save a lot of money during various research approaches.

Dr. Reisch believes that the next generation of biostorage solutions will be much more compact; operate at –20°C and –80°C at the same time, if needed; minimize thaw/freeze cycles; save energy costs compared to standard freezers; and be scalable, even if the customer wants to expand the store after several years.

The controlled environment provides higher sample-quality that will reduce costs during the research phases of drug development and enable biobanking on a worldwide level because experiments will be more accurate and reproducible, Dr. Reisch concludes.

“With the shift from HTS to HCS and consequentially the focus on studies from biochemical assays to cell-based studies, libraries for storing biological material are in increasing demand,” notes Stefan Betz, product manager at Thermo Fisher Scientific (www.thermofisher.com). “One area of focus is the identification of targets involved in a disease pathway.”

Thermo Fischer’s BioBank™ stores biological samples at –80°C for applications such as cell-based assays; bacterial clones; and protein, DNA, and RNA libraries. BioBank holds thousands of samples in a range of configurations. A rack-design system enables the location of the robotics in an ambient temperature, isolated from the ultralow temperature that can cause mechanical failures. Its CO2 backup protects samples for up to 12 hours during power outages.

Ensuring sample integrity during long-term storage is a critical element of success for many research organizations, Betz says. Because BioBank is a stand-alone unit, sample storage/retrieval and preventative maintenance do not compromise sample integrity during long-term storage.

At the Harvard Medical School Institute for Proteomics, BioBank enables the distribution of small and large sets of clones to researchers with custom orders. A tube-picking system allows for an experiment-driven array of plasmid clones in a microtiter plate format that will enable customers to use the arrayed samples directly in their experiment.

“BioBank enables drug discovery processes that use sensitive, biological samples to be automated in a flexible, high-throughput capacity,” Betz states. “Samples can be easily selected from the -80° environment, thawed under controlled conditions, and presented to external automation for processing in the drug discovery process of interest. This step in itself improves the quality of the data obtained in subsequent steps while avoiding sample degradation.”

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