Biomolecule Production Core

The overall objective of this Core is to provide the necessary infrastructure for the expression and purification of biologic molecules including antigens, proteins, carbohydrates, nucleic acids and other biologics from the Risk Group 4 and 3 agents for investigators associated with the NEIDL. The design of the NEIDL incorporates both BSL-3 and BSL-4 Biomolecule Production Core laboratories. We believe that this design maximizes the efficiency of research that will be performed under high containment in the Institute by minimizing the extensive downtime that would be required for conversion of a single flexible laboratory module. Moreover, this design allows for concomitant use of both high containment laboratories, thereby increasing by more than 2-fold the total work flow in this Core. For example, at the same time Kyasanur Forest Disease virus can be propagated for challenge studies in the BSL-4 Biomolecule Production Core laboratory, Francisella tularensis may be grown for the extraction and purification of capsule to be used in an experimental conjugate vaccine in the BSL-3 Biomolecule Production Core laboratory. From an operational perspective, not all personnel trained for work in the BSL-3 Biomolecule Production Core will necessarily have to be certified to work under BSL-4 containment. It is clear that the Standard Operating Procedures (SOPs) governing the growth and purification of proteins and carbohydrates from BSL-3 bacterial pathogens (e.g., Francisella tularensis, Yersinia pestis, multi-resistant Mycobacterium tuberculosis); versus BSL-4, the growth and extraction of proteins and nucleic acids from the BSL-4 viral pathogens (e.g., Kyasanur Forest Disease, Ebola, Marburg, Lassa), as well as decontamination procedures, and operational plans will be different and appropriate for each level of containment.

Functions and Services

The Biomolecule Production Core at BSL-4 will have dedicated facilities and production capabilities to grow Risk Group 4 viruses and isolate biologic molecules (e.g., antigens, proteins, nucleic acids, etc.) of interest under BSL-4 containment. The workscope for the BSL-4 facility will be strictly governed by the NIH Guidelines for Recombinant DNA Research, as well as the City of Boston regulations. The Core will establish a collection of BSL-4 agents (e.g. filoviruses, arenaviruses, bunyaviruses, henipaviruses) and produce and purify sufficient quantities of biologic molecules of interest to investigators affiliated with the NEIDL in order to conduct pre-clinical in vitro and animal evaluations. When preclinical evaluation warrants further development of a particular biologic molecule, the core will work closely with the expertise of the Massachusetts Biological Laboratories to effect technology transfer and assist in efforts to optimize methods for biologic molecule production of those “products” under a lower level of biologic containment in anticipation of transfer of production to this GMP facility enabling manufacture of clinical grade materials for phase I/II., phase II, and phase III clinical evaluation. In order to aid downstream purification for initial testing, the Core will, as appropriate make use of affinity tagged systems. It is recognized that while the use of affinity tags are attractive for initial purification, the inclusion of tags presents a challenge for subsequent Food and Drug Administration approval. For this reason, in those instances where an individual protein shows pre-clinical promise, the Core will develop expression and purification systems that do not employ affinity tags. The Core will also provide inactivated material from various pathogens that can be used as antigens for antibody production. Prior to production, Master Stocks and Seed Stocks will be prepared from single isolated viral plaque. Seed Stocks will be characterized according to the Points to Consider from the Food and Drug Administration, including genotyping to provide a database for positive identification of strains that may be used in a terrorist event. The Core will develop standard operating procedures (SOPs) for the growth and expansion of all biologic cultures and their purification of all biologics. The Core will also establish methods and procedures for maintaining appropriate levels of security and controlling inventory of the biologic agents acquired by and used by the NEIDL Investigators. For example, single vials of each Master and Seed Stock will be individually labeled and stored in a locked - 80°C freezer. Accurate and up to date inventories will be maintained, and physical inventories will be periodically conducted to ensure that records are accurate. Signing out of materials from the Core will require signatures of two Core scientists, and receipt signature of the requesting NEIDL Project Director / Scientist.

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