Biology: Crystallographic Studies of Biohazards (Biosafety Levels 2 & 3)


BioCARS is a National User Facility for macromolecular crystallography. It provides state-of-the art facilities and scientific and technical support for standard X-ray diffraction techniques (such as MAD). But the facility is also designed for two special tasks that set it apart from other centers for macromolecular crystallography at the APS and nationwide. The BioCARS insertion device beamline 14-ID can serve as both monochromatic and polychromatic X-ray source. The polychromatic capability is essential for conducting time-resolved X-ray diffraction experiments that utilize the Laue technique. These experiments result in molecular movies depicting biologically important macromolecules as they perform their function. The other special feature of the BioCARS facility is that the experimental stations are embedded in a Biosafety Level 3 (BSL-3) containment, unique at the APS and nationwide. This permits safe research with biohazardous materials classified as Biosafety Level 2 or 3 agents, such as human, animal or plant viruses and toxins.

Agents classified as BSL-2 involve a broad range of indigenous moderate-risk agents that are present in the community and associated with human disease of varying severity. Immunization or treatment for these agents is available. Examples include: measles virus, Salmonellae and Hepatitis B virus.

Agents classified as BSL-3 are indigenous or exotic agents that may cause serious and potentially lethal human diseases as a result of exposure by the inhalation route. Examples of BSL-3 agents include M. tuberculosis, the St. Louis encephalitis virus and Coxiella burnetii. The guidelines for BSL-2 and BSL-3 facilities, described in the "Biosafety in Microbiological and Biomedical Laboratories" CDC/NIH publication (Fourth Edition, May 1999), require special safety equipment as well as a special facility design for handling BSL-2 and BSL-3 materials. All of these requirements are fulfilled by the BioCARS facility, originally designed by Wilfried Schildkamp and recently upgraded for BSL-3 operation in a joint effort by BioCARS staff (led by Vukica Srajer and Reinahrd Pahl) and APS (led by John Sidarous and Marvin Kirshenbaum). The facility was approved for BSL-3 operation on November 2, 2004 by ANL Institutional Biosafety Committee. The first BSL-3 experiment was conduced on November 12, 2004.

Manipulation of all BSL-3 agents is done in a Class II Biosafety Cabinet that serves as the primary protection barrier. The facility is physically separated from the access corridors and is maintained under negative pressure (with respect to the APS experimental hall) while BSL-2 and BSL-3 experiments under way. Air flows from the experimental hall into the facility and is exhausted from the facility via a HEPA filter out of the APS building. Mandatory Standard Operating Procedures for BioCARS users and staff have been developed and describe in detail the set-up and operation of the facility in the BSL-2/3 mode, rules for conducting BSL-2/3 experiments, the response to incidents and emergencies as well as the clean up procedures and safe disposal of any waste generated during such experiments.

The primary scientific and practical expertise for the work with BSL-2/3 agents resides with the BioCARS users. Many virus crystallographers nationwide already use the BioCARS facility for their research projects. In this expanding field of national importance, BioCARS is in a position to play an important role as the only synchrotron-based X-ray diffraction laboratory that can be used for safe study of such samples.