An Assessment of the National Institute of Standards and Technology Material Measurement Laboratory: Fiscal Year 2014 (2015)

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Biomolecular Measurement Division

The objective of the Biomolecular Measurement Division (BMD) is to develop measurement technology, standards, and data to determine the composition, structure, quantity, and function of biomolecules. The work focuses on bioprocessing, applied genetics, mass spectrometry, biomolecular measurement, biomolecular (protein) structure and function, and bio-analytical science. These areas are identified using terminology that is consistent with the terminology used for research areas in systems biology and molecular biology. The actual work being carried out by the BMD appears to be more focused than what is indicated by the groups’ titles. Specifically, the BMD, which has 11 staff in its headquarters office and 100 others elsewhere (including 30-35 guest researchers), is divided into projects conducted by five groups.

The Mass Spectrometry Data Center develops mass spectral libraries for small molecules, metabolites, peptides, glycans, and proteins; the Applied Genetics Group develops standards and technology for DNA testing to aid human identification efforts and to support biometric, law enforcement, and clinical applications using genetic information; the Bioanalytical Science Group develops protein-based standard reference materials such as C-reactive protein (CRP) and bovine serum albumin (BSA); the Bioprocess Measurements Group develops measurements for counting and sizing biological particles (protein aggregation and preparation of protein particle surrogate materials as SRMs); and the Biomolecular Structure and Function Group performs work in crystallography, neutron scattering, and nuclear magnetic resonance (NMR) to determine the structure of biomolecules (including a monoclonal antibody standard).

The five groups carry out their work on the main campus and at the Institute for Bioscience and Biotechnology Research (IBBR) at Rockville, Maryland. These groups appear to have distinct goals, but at the same time they carry out work on complementary research topics and technical themes directed at the forensics and diagnostics of nucleic acids (DNA), peptides, proteins, and macromolecules in addition to smaller molecules. Characterization of chemical molecules and peptides is the major mission of the Mass Spectrometry Data Center. A significant capability of the BMD is at the IBBR, which carries out work on proteins and macromolecular structures with initiatives in biopharmaceutical measurement and engineering. This work is centered on the facilities in Rockville, including a 900 MHz NMR spectroscopy system housed in the W.M. Keck Structural Biology Laboratories. In addition to fundamental research related to a range of proteins, nucleic acids, and nucleic-acid protein complexes, NMR methods are also used to study protein folding, stability, and interactions in solution in order to develop models, standards, and databases.

The number of people associated with the groups is given in parentheses: Mass Spectrometry Data Center (32), Bioprocess Measurements Group (22), Biomolecular Structure and Function Group (20), Bioanalytical Science Group (13), Applied Genetics Group (12), and the Biomolecular Measurement Division headquarters (11).

Thirty-five percent of the division’s researchers and employees consist of guest researchers and 65 percent are federal employees. The budget of the BMD is $23.3 million (fiscal year 2014) with $5.47 million of the $8.86 million biomanufacturing initiative being awarded to the BMD. There are three

primary products of the BMD: measurement science and protocols disseminated through scientific publications, standard reference materials and reference data, and spectral libraries.

ASSESSMENT OF TECHNICAL PROGRAMS

Accomplishments

While the biomolecular component of the BMD suggests that the activities of this division focus on measurement science, standards, and data and technology for biomolecules and biomacromolecules. A major component of this division’s work also addresses reference data and spectral libraries for smaller compounds (including biologically relevant low-molecular-weight metabolites and peptides) through the Mass Spectrometry Data Center. The center’s work is a major component of the BMD’s overall effort and has national and international impact for the identification of chemical entities through their mass spectral fingerprints. The center is responsible for NIST-evaluated spectra, together with validated software for spectral matching packaged in a user-compatible visual interface. These products are widely integrated into instrument vendor software systems. The reference database is the world’s most widely used mass spectral library and is delivered, together with commercial instruments, through the companies that produce, market, and service mass spectrometers. More than 5,000 new installations of these reference data and software occur each year. The mass spectral library is an important component of standard reference data sales by the MML. The MML accounts for 98 percent of standard reference data sales in NIST. In 2013, 79 percent of MML sales were derived from the mass spectral library.

The work on measurement science, mass spectrometry data, and protocols for forensics involving DNA are major and continuing contributors to the impact of the BMD. Many BMD activities are oriented to understanding the impact of alterations in protein structure that indicate changes in identity, purity, or function, thereby creating the knowledge that will be critical for biomanufacturing or health regulation. This effort involves significant exploratory research and is defining the problems that need to be addressed. The donation of a significant quantity of a humanized monoclonal antibody (mAb) by Medimmune, LLC (now a division of AstraZeneca) has provided material for a wide variety of development efforts focused on protein metrics and models that describe effects of the protein’s environment (pH, ionic strength, other proteins, etc.) on structure and function. This focus involves a major effort not only in the BMD, but across the MML, and fosters internal and external collaborations. Discussions with researchers at the IBBR suggested that this mAb is being used as a reference against which the characterization of other proteins may be compared as the BMD advances its capabilities in protein characterization.

The Bioprocess Measurements Group combines expertise in instrumentation science, nanomaterials, and chemical and physical characterization of surfaces, sensor materials, and biological molecules to address measurement needs relevant to biological industrial processes.

The Biomolecular Structure and Function Group is located primarily on the main Gaithersburg campus, with another facility near Rockville and a University of Maryland facility nearby. These locations enhance the group’s collaborations external to the MML, especially with the University of Maryland, industry, and—separately—with the FDA. This group is focused on measurement of disease pathways, vaccines, protein therapeutics, and biomarkers for diagnostics. The studies on proteins in the new 900 MHz NMR, compared to those produced by more common instruments, will inform more efficient use of the latter for bioprocess analysis. The studies expand the study of proteins to include RNA, lipids, and glycans. Cutting-edge instrumentation is used to develop measurement technology to extract information on molecular function, structure, and responses to molecular interactions and changes in the environment (e.g., temperature, humidity, pH, and sample matrix). These types of data are critical for the bioprocessing industry and biotherapeutic applications.

The Bioanalytical Science Group is developing reference materials based primarily on proteins, including reference standards for bovine serum albumin, C-reactive protein, and an IgG mAb. The

technologies used to characterize these materials are state of the art, and the staff are very aware of the need to distinguish properties relevant to function in addition to those indicative of purity and structure. The data obtained from characterization of the NIST mAb will be published in an American Chemical Society book by the end of 2014 and will be the most exhaustive analysis of a single protein performed to date, with more than 60 collaborating partners involved.

The Applied Genetics Group validates instruments and methods used for forensic genetics. It provides DNA profiling reference materials that are used to ensure that forensic laboratories are generating accurate and reliable results. The group has produced a DNA reference material for clinical diagnostic laboratories to ensure the accuracy and comparability of test results for Huntington’s disease. It is expanding this program into other clinical areas. An extensive sequencing program is testing methods and instruments for rapid polymerase chain reaction (PCR) technologies and rapid gene sequencing. This program has produced a variety of standard reference materials, methods, and data.

Review of the accomplishments of the five groups shows a range of valuable products for clinical diagnostics; genetics; viral genetics; pharmaceutical products; environmental, food, and forensic analysis; biosensor development; structural characterization of large molecules (particularly proteins); and the previously mentioned mass spectrometry database. The achievements are significant and impact the pharmaceutical, clinical diagnostic, chemical, law enforcement, and biopharmaceutical sectors (Table 3.1). Some of the more recently initiated work in structure/function studies for protein therapeutics and biologics needs more time to develop, given the rapidly changing character of the biotechnology industry and the demands for new products that address human diseases in a cost-effective manner. Achievements and impact will likely evolve as the capabilities and focus of IBBR activities continue to develop and data from this facility become available, at first internally in the MML and then more widely to the biopharmaceutical industry.

Opportunities and Challenges

The research on protein structure and modifications that impact function is needed, but the challenge is to focus the research so that it complements related work supported by industry as well as the $25+ billion budget of the National Institutes of Health (NIH). There is recognition at the division and group levels that such interactions are important, but a specific approach to how the needed connections will be made and used to inform the work of the BMD is less clear.

TABLE 3.1 Biomolecular Measurement Division: Summary of Activities and Achievements

NOTE: LC/MS, liquid chromatography/mass spectrometry.

As an example of one standards development effort for biomanufacturing, the Bioprocess Measurements Group is examining measurement of protein particles and aggregate formation, leading to the preparation of a nanostructured particle that has a shape similar to that of a protein aggregate. While this is an impressive technical achievement, it is not clear that the scientists have given thought to the other factors that need to be considered, because shape alone is not sufficient to define a protein aggregate. Other factors that need to be considered include amino acid sequence, cross-links formed, solution pH, and temperature history.

A significant issue that the BMD would like to pursue is to develop tests for protein potency. How does one relate physicochemical characteristics to protein functionality? This is a very important question whose answer would involve collaboration between the BMD and the Biosystems and Biomaterials Division of MML, and probably some of the other divisions. If the BMD could discover some revealing physical metrics that are reliable indicators of function, expensive tests using cell culture and whole animal systems could be drastically reduced.

The BMD has the opportunity to develop cross-disciplinary culture and to pursue complex issues faced by the growing biomanufacturing sector in the United States. This sector may expect process-specific standard reference data and materials, while the BMD will need to be able to prepare materials that have a broader utility across the industry. If this is the case, strategic allocation of capabilities and resources within the BMD, and more broadly the MML, will need to be achieved in the coming years. A team approach with specific or allocated tasks for each team member will need to be set forth and managed.

Overall Assessment of Technical Programs

Overall, the BMD program includes strong individual efforts, but the complexity of biomolecules requires that a more integrated approach be defined. The need to understand information relevant to function as well as structure needs to stay in the forefront as future priorities for programs are established. An overall assessment of the BMD’s technical programs shows programs consistent with the mission of NIST and the MML. The BMD is addressing biotechnology for forensics and biomanufacturing and is part of the initiative in big data.

The BMD groups show strong individual efforts, while at the same time providing complementary expertise among the programs pursued by the five groups (refer to Table 3.1). All of the groups show productivity in publishing (roughly one paper per person per year) and align with the BMD’s goal of disseminating measurement science and protocols through scientific publications. In addition, the groups also provide either reference data or reference standards to the industry, as summarized in Table 3.2.

At the same time, the complexity of protein structure and function and the classification of some of the proteins in terms of their production process (rather than just their composition) add complexity to the approaches that need to be developed. Unlike chemical or peptide reference data or samples, protein data need to reflect differences in protein glycosylation patterns, protein cross-linking, refolding patterns, and post translation modification—that is, changes that occur after the cell expresses or releases the newly formed protein.

The technical programs, and particularly the intragroup dynamics, appear to be good. However, the goals and the products to be generated as a consequence of these goals still need to be defined in some cases. In the case of chemical molecules, the goals and framework for achieving the goals are clear, and the instrumentation and analytical methods needed to carry out the measurements are available and readily carried out.

TABLE 3.2 Standard Reference Materials Prepared by the BMD

The reorganization of the MML was carried out to achieve more formally managed cross-division technical programs and to address cultural, organizational, and management barriers to staff interactions and collaboration across divisions. While strong interdivision collaboration needs to be proactively encouraged, cooperation and collaboration between groups within a division may also need to be encouraged.

The BMD was formed as part of the MML reorganization, and the division is working on enhancing collaborations and cooperative work within the division. A division seminar series was

mentioned as a tool to help people get acquainted with the work of others both on an intra- and interdivisional basis. While this is a start, a first impression is that further development of intra-divisional work, particularly between Gaithersburg and Rockville (IBBR), is needed. While the biomanufacturing program has assisted in cross-division collaboration, this cooperation needs to be reinforced and further developed.

Biologic drugs are both a U.S. economic driver and a health care cost issue and may benefit from the establishment of standards. The recent focus of the BMD on biologics, particularly through the IBBR, is appropriate but also complex, in that the molecules to be studied, and for which reference data and/or standards are needed, are also very complex and manufacturer-specific. However, if the BMD is to successfully address this challenge, close cooperation between the Bioanalytical Science, Bioprocess Measurements, Applied Genetics, and Biomolecular Structure and Function groups will be needed to address standards at each step of the research, development, and, ultimately, manufacturing processes: (1) identifying a protein therapeutic candidate; (2) cloning and development of a host organism, (3) cell culture and protein production, (4) protein purification, and (5) formulation and fill.

These steps are part of a complex manufacturing process, where each step may impact the characteristics of the biomacromolecules. The BMD has recognized the inherent challenges, but the groups need to work together to identify where concerted efforts would tap into the unique capabilities of the BMD and the MML to make a difference in reducing costs of biologic drugs through standards that only the MML could provide.

The workshops that the BMD has organized and participated in have addressed topics that will assist in developing strategies for entering new areas of biomanufacturing where standards are needed. Examples of these workshops include a biomanufacturing summit, applications of mass spectrometry for biopharmaceutical manufacture and characterizing biopharmaceuticals in a regulatory setting, and next-generation characterization tools for therapeutic proteins. Overall, these programs and activities represent excellent first steps in defining the role of the BMD in developing standards and meeting the goals of the MML for the next-generation bio-based products.

PORTFOLIO OF SCIENTIFIC EXPERTISE

The scientific staff have made significant accomplishments. There is significant variability in publication output, from easy-to-measure to the more difficult to quantify information and standards provided to industry. Overall the output is respectable when compared to the measure of papers that are published by academic counterparts in related fields. These numbers could be higher, but many of the staff are engaged primarily in exploratory research. However, the value of the work lies in its development of clearly defined standards. One such standard might specify, for example, the forensic accuracy of commercial gene typing instruments, which are much more complex than, for example, the concentration of BSA in solution. While both are important, defining the reliability of an instrument or protocol used in courtroom proceedings is a task that the MML would seem uniquely qualified to do.

The opportunities and challenges in terms of scientific expertise are to integrate the historic strengths of the MML, that is, standard reference materials, with the technologies used to characterize and produce them. This is particularly important in the biotechnology industry, because the products are not as well defined chemically or physically as, for instance, the molecules encountered in metallurgy, organic chemicals, small molecules, pesticides, and other more discrete chemical entities with which the BMD has significant experience. Consequently, a directed effort will be needed to carefully foster expertise relevant to the development of standard measurements for characterization of biomolecules (an example would be the skill required to characterize a monoclonal antibody using 900 MHz NMR).

The portfolio of scientific expertise at the division level has individuals who are outstanding scientists in their area and who work well as members of teams defined by discipline or investigating a specific target material or molecule. As the MML moves into the biotechnology and bioproducts areas, the BMD needs to define the types of measurements and standards the industry will need. These could be

diverse, ranging from standards for improving efficiency of manufacturing technologies for biomolecules used in pharmaceuticals, to protocols for improving diagnostics of human or animal diseases or identifying biomarkers indicative of disease. There is the potential for considerable overlap of the activities of the BMD with the activities of other agencies, so goals need to be selected that focus on work that is synergistic with the goals of those other agencies.

ADEQUACY OF FACILITIES, EQUIPMENT, AND HUMAN RESOURCES

The BMD reported many significant accomplishments. The successful projects varied from developing instruments and defining protocols for forensic genomics for personal identification to characterization of complex proteins that define some types of biopharmaceutical molecules. The developments with respect to characterization of proteins may in some cases lag the field with respect to knowledge and protocols already developed in the industry but may in other cases be useful for identifying appropriate measurement techniques and instruments that could give useful data in the absence of extensive and expensive facilities like those at the BMD. The group with the most internationally recognized accomplishments is the one that operates in the area of mass spectrometry—this mature team is large and highly focused.

The BMD has tremendous intellectual capital, state-of-the-art facilities, and a mandate to develop standards and protocols for materials that are likely to emerge from the continuing and rapid development of biomolecules, including biopharmaceuticals, biopolymers, and specialty bioproducts used as precursors in biochemical manufacture. The challenge will be to apply the limited resources available to the BMD wisely and to target areas that cut across different industries or companies in a way that benefits the industry and, at the same time, is accepted by individual companies as part of their manufacturing technology and regulatory ecosystem. This will require a deep understanding by the BMD of the manufacturing processes that these companies utilize and the ability to propose reference materials and information that would assist them in developing their manufacturing processes. Consequently, the staffing of the BMD needs to reflect this mission, and the division may need to increase their exposure to problems of biomanufacturing in order to carry out targeted work in their mission of biomolecular measurement.

The BMD has excellent facilities and human resources that could have a major impact if they are directed toward specific targets and industry needs that transcend the capabilities of large or small individual companies to address. A survey of these needs by an organization such as the Pharmaceutical Manufacturers’ Association (PMA) or one of the committees of the Biotechnology Industry Organization (BIO) may be useful.

DISSEMINATION OF OUTPUTS

In addition to the mass spectrometry database and the widely used albumin standard, the outputs of the BMD include DNA standards, biomarkers, and forensic training.¹ In all these cases, the information that is bundled with the standard reference materials has made a major impact on the industry and the ability to manufacture products (even if by different processes) that have common characteristics, identifiable qualities, and safety. This is a key to the contributions made by the BMD and also to the recognition of the value of the MML in impacting the industry in many but subtle ways. It is important to maintain this combination of data, information, and reference materials.

The provision of standard reference materials, vetted results on new instruments, and assessment of data validity are needed to ensure that industry and law enforcement agencies are able to produce reliable information. The challenge is in developing efficient and effective approaches to obtaining a deep

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¹ NIST Material Measurement Laboratory, Turning Ideas Into Innovations, Gaithersburg, Md., August 2012.

understanding of biomanufacturing processes, which are often less well defined and at much earlier phases than the mature industries. This knowledge is needed to ensure that dissemination of outputs is carried out in a useful, timely, and accurate manner and that the MML is able to increase the efficiency of innovation and manufacture of bio-based products. The dissemination of web-based products has not realized its potential, and there is plenty of opportunity for improvement.

The analytical focus on biomolecules is yielding important information and opportunities to develop standards. The next step, extraction of knowledge from the data, needs increased attention in terms of how to do it, how to understand it, and how to share it.

FINDINGS AND RECOMMENDATIONS

The recommendations below relate principally to how the goals and metrics of performance are articulated, with emphasis on the importance of maintaining many of the positive aspects of the BMD that make it a unique and valuable resource to the industry.

A BMD strategy integrated with an overall MML strategic plan is needed to clearly map the pathway from current BMD strengths to the development of excellence in applying standard metrics to biomolecules.

Data acquisition is only the first step in providing standards for the biotechnology area. Developing a focused and integrated plan on data informatics is particularly important for BMD standards efforts.

The BMD needs a much deeper understanding of biomanufacturing processes, which are often less well defined and in much earlier phases than the mature industries that MML has historically addressed over the past 20 years. This understanding is needed to ensure that appropriate research targets are selected at the MML and that generation and dissemination of outputs to the biomanufacturing industry are carried out in a useful, timely, accurate, and relevant manner.

The BMD is struggling to derive the knowledge to underpin the selection of metrics critical for biomanufactured products. Developing such an understanding is a difficult and very important challenge. The BMD needs more collaborative interactions to achieve this knowledge.

Personnel management at the BMD appears uneven and handled with variable effectiveness, resulting in very different expectations, especially among junior staff. Mentoring of postdoctoral researchers tends to be excellent, but mentoring of junior permanent staff is very uneven. Understanding of technical goals, especially those of cross-divisional projects, is limited in some cases. With the exception of their frustration due to contracting and technology transfer administration, most staff are very happy about the working environment. However, it is important that the BMD management communicate the technical strategic plan to all its staff, make supervisory processes transparent, and remove, to the extent possible, unnecessary roadblocks to purchasing and collaboration. These actions are needed if the excellent morale, can-do attitude, and commitment to excellence in measurement results are to be maintained within the BMD.