DNA-impersonator gets ISO Class 8 hideout

by Mark A. DeSorbo

Biotech R&D firm more than doubles in size, sets its sights on cGMP

Viruses may run rampant, but they cannot hide from technologies like those of a Bedford, MA-based biotech firm's DNA-impersonator, a blood culture canvassing agent that shines a fluorescent spotlight on trespassing microorganisms.

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Synthesizing peptide nucleic acid (PNA) to mimic DNA and meet the demands of the clinical diagnostics and life sciences markets has been the focus of Boston Probes' research and development since it opened its doors six years ago, says Tish Creasey, director of operations.

With PNA being an emerging technology, research and development efforts have intensified at Boston Probes, and Creasey says the firm's goal is to apply for current good manufacturing practice (cGMP) certification from the U.S. Food and Drug Administration (FDA) by the spring of 2002.

Boston Probes’ new ISO Class 8 (Class 100,000) cleanrooms may seem vacant, but the biotech firm was still getting settled at the time of this report. Along with a separate mechanical room, the cleanroom suite has six separate environments that share a common hallway.
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Moreover, with an expiring lease and limited space for growth, Boston Probes needed to find a new home. “The lease was up and the facility we were in before limited us to a maximum of 6,000 square feet,” adds Creasey, who worked with contractors in the renovation of Boston Probes' new facility. “There was just no room for expansion, and space was essential for us.”

Leaving the town of Bedford, a suburb of Lowell and Boston, was not an option either, for it was the best geographic location for Boston Probes' employees. The company luckily found a 30,000-square-foot building nearby in the same town.

At the time of this report, Boston Probes had just started moving into the facility that had 20,000 square feet renovated into offices, laboratories and cleanrooms. Creasey says 1,500 square feet has been designated as ISO Class 8 (Class 100,000) cleanroom space. The remaining 10,000 square feet remains dark, but on hand for anticipated growth.

Cutting and stabilizing
Leading the project was J.M Coull Inc., a Concord, MA-based construction-management firm versed in cleanroom design and build projects.

“This was an existing building that we gutted and renovated,” says Andrew A. Coull, executive vice president. “The project took about four months to complete, and the bulk of it was completed in August.”

Finishing touches were being made at press time to the biological level-2 hazardous material storage room. Coull also reports that Boston Probes had a targeted move-in date. “They wanted to start moving in on August 10 and they began moving in on August 10,” he adds.

Neither Coull or Creasey would provide details about the cost of the cleanrooms or the project's budget, but Coull adds that “the project was within budget and we worked the design to fit that budget.”

Coull and Creasey, however, did say that they had a pre-construction plan, which outlined layout, programming, budgeting and scheduling. “We reviewed these areas before we started building so we knew how much it would cost and how long it would take,” he adds. “We worked very closely together.”

That planning precluded painstaking problems as well.

While coordinating timely town inspections posed minor difficulties, the real challenges came when construction crews needed to build additional supports for the HVAC system on the existing roof. Significant undercutting of existing concrete slab also had to be done to install plumbing for labs and cleanrooms.

“We had to build a platform on top of the roof on the existing structural column to support the HVAC systems for the cleanrooms and the laboratories,” Coull says. “We did not factor that in beforehand. We knew we had to deal with it, but we did not have the solution until we were able to study the existing structure and devise the solution.”

The most expensive challenge was cutting concrete and digging into the existing foundation to install piping for laboratory and cleanroom sinks. “The expensive thing is to do the cutting and then the most challenging is to fit the plumbing in to the shortest distance between two points,” Coull says.

Some sinks just could not be moved, but there were some that could and it saved 20 feet of cutting, piping and backfill. “Instances like that resulted in significant cost reductions,” Coull adds.

Exposing bacteria
When it was all said and done, Boston Probes had an ISO Class 8 (Class 100,000) cleanroom suite of six 12-by-15-foot rooms, with a common hallway.

“It is a dramatic improvement over what they had previously,” Coull says.

Cleanrooms are serviced with a de-ionized water system from Millipore Corp. (Bedford, MA). Controlled areas breathe through rooftop air-handlers and an HVAC system from Trane Co. (Lacrosse, WI) and Mac 10 HEPA filters from Enviroco Corp. (Albuquerque, NM).

The HVAC system was designed and installed by Thermo Engineering (Worcester, MA), notes Daniel Mahoney, vice president of J. M Coull. “It controls air flow, temperature, moisture, relative humidity and pressurization in individual cleanrooms,” Mahoney adds.

A blood culture sample that has been treated with PNA. The PNA attached and fluoresces to the bacteria it has been geared to detect.
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While there is some make-up, air is re-circulated and the temperature is maintained at 68 degrees plus or minus 2 degrees Fahrenheit, with a humidity level of 45, plus or minus 5.

Cleanroom personnel, Creasey says, wear protective garments, such as lab coats, gloves and eye protection, but they are not required to wear shoe covers. “It's a relatively small space. It's an organic chemistry environment, so there are [fume] hoods,” she adds. Personnel also wear protective garments when working in the facility's negatively pressurized biological level 2 room.

With new labs and cleanrooms, officials at Boston Probes are now concentrating on getting their proprietary technology to market, says Henrik Stender, director of microbiology at Boston Probes.

By synthesizing PNA, Stender explains that specific probes can be developed to detect almost any microorganism. “You can make specific probes for E. coli, tuberculosis, HIV, Candida yeast,” he says “You can also use it to target genetic material to identify certain chromosomes.”

It works by taking samples from blood cultures that have been incubated and show signs of growth, Stender explains. The sample is gram stained to determine if gram-positive [bacterial] cocci are present. PNA is then applied and incubated for 90 minutes.

“In that time, PNA probes enter the cells and once it detects what it was geared to find, the PNA shines,” he says. “When you look inside the microscope you can actually see the individual cells fluoresce; the microorganisms are lit up by the probes.”

Once the microorganism is identified, the correct therapy can then be administered to patients, says Creasey. “PNA is geared to go after highly structured DNA sequences or ribosomal RNA,” she adds. “Our technology can identify the type of virus so that patients can be treated effectively.”

Key facts
Size of facility: 30,000 square feet, 1,500 of which is designated ISO Class 8 (Class 100,000) cleanroom space.

Purpose of facilities: To develop peptide nucleic acid as a DNA mimic for the clinical diagnosis and life science markets.

Designer(s): J.M. Coull Inc. (Concord, MA) designed and renovated the facility, while the HVAC system was designed and installed by Thermo Engineering (Worcester, MA).


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