Bridging the gap

Figures & data

Figure 1 Early stage development of a therapeutic mAb spanning antigen identification through discovery, expression and purification. This preclinical stage represents an iterative process in which results from testing and evaluation feed back on the molecular elements of the mAb.

Figure 2 A client-based analysis of client projects conducted by the AIBN Biologics Facility over the past three years. (A) Contract distribution based on phase of development. (B) Total output percentage based on the litres of culture used per project category.

Table 1 Top selling drug products worldwide

Table 2 Academic and government funded facilities for early stage development of biologics

Facility and Location	Brief Description	Contact
Biopharmaceutical Development Program (BDP). SAIC-Frederick, Inc.,	The BDP is a cGMP-compliant facility that occupies/manages approximately 48,000 square feet of processing area, most of which is environmentally controlled, on the NCI-Frederick campus, and maintains a Type 5 Facility DMF with the FDA. The BDP produces clinical-grade biopharmaceuticals, including monoclonal antibodies, recombinant proteins, immunoconjugates, peptide and DNA vaccines, viruses and other biologicals required by NCI for Phase 1 and Phase 2 clinical studies and proof of- principle evaluations. To accomplish this mission, they maintain expertise in the areas of production, purification, analysis, vialing and FDA regulatory compliance of biologicals. The facility has mammalian cell reactors up to 1,000 L (single use and STR), full downstream capabilities (including automated skids for cGMP purification) and automated filling and labeling systems. In addition the facility provides extensive process analytics and quality control assays and well as guiding regulatory affairs and quality assurance eg CMC, IND, Drug master files etc.	John A. Gilly, Ph.D. (Contractor) Associate Director, National Cancer Institute at Frederick P.O. Box B Frederick, MD 21702-1201 USA Phone: [phone] E-mail: [e-mail] Web: wwwbdp.ncifcrf.gov
Center for Biomedicine and Genetics (CBG)	CBG is designed specifically to accommodate small scale (bench-top) production of GMP-grade biologics, viral and nonviral gene therapeutics and modified patient cell populations to support phase I and II clinical trials at City of Hope. Adherence to cGMP is essential in ensuring the quality and integrity of manufactured biologics. The facility is a 20,000 square foot California-licensed multiproduct manufacturing facility with 12 production and purification rooms. In addition the facility has 57 staff and works with a diverse range of projects. The facility also has a Class 100/100 aseptic fill suite.	Larry A. Couture, Ph.D. Senior Vice President, Center for Applied Technology Development City of Hope 1500 East Duarte Road Duarte, CA 91010 USA Ph: [phone] E-mail: [e-mail] Web: www.cityofhope.org/research/support/center-for-applied-technologydevelopment/center-for-biomedicine-andgenetics/Pages/default.aspx
Biotherapeutics Development Unit (BDU)	The BDU is responsible for the manufacture of clinical grade biological products, and is part of Cancer Research UK's Drug Development Office. The unit is run to cGMP and has a license issued by the Medicines and Healthcare Products Regulatory Authority for the manufacture of investigational medicinal products. The BDU has the capability to manufacture plasmid DNA, monoclonal antibodies and recombinant proteins. The development department provides an efficient means of scaling up the manufacture of research products. The QC section carries out all the necessary quality control testing to ensure the quality, safety and efficacy of the product, and has the ability to develop new and existing assays to meet customers' needs.	Gillian Lewis Cancer Research UK Clare Hall Laboratories Blanche Lane South Mimms Herts. EN6 3LD United Kingdom Ph: [phone] E-mail: [e-mail] Web: science.cancerresearchuk.org/sci/bdu
Children's GMP, LLC (St. Jude's)	Children's GMP, LLC is a wholly subsidiary of St. Jude Children's Research Hospital. Their mission to provide first in human clinical trial materials under cGMP regulations exclusively for St. Jude investigators and their collaborators. Focused on manufactured monoclonal antibodies, gene therapy vectors, protein and viral vaccines and therapeutic proteins (from mammalian cells and bacteria). The facility is ∼67,000 square feet and includes approximately 2,000 square feet of GMP suite and laboratory support space. This includes 12 GMP production rooms, including 2 BSL-3 suites, separate cell banking suite and a separate fill/finish suite. In addition the facility offers a dedicated Quality Control laboratory and raw material and finished product storage rooms. The facility has 5, 30 and 100 L STR dual-purpose bioreactors/fermenters as well has two 20/50 WAVE type. For downstream purification the facility uses an ÄKTA Prime, 3 × ÄTKA Explorer 100s and an ÄKTA Pilot as well as a range of tangential flow filtration systems and a validated a semi-automated fill and finish system (Millipore Acerta). The facility has a staff of 9 production personnel, 5 QC, 4 QA, 3 validations, 3 Materials Management staff.	James Allay. St. Jude Children's Research Hospital 262 Danny Thomas Place Mail Stop 920 Memphis, TN 38105-2794 USA Ph: [phone] E-mail: [e-mail] Web: www.stjude.org
Bioprocess Scale-Up Facility (BSF)	The BSF is the Biotechnology Research and Education Program's modern bioprocessing laboratory dedicated to the development and scale-up of biotechnology products and processes. With over 1,000 fermentations conducted since 1998 alone, the BSF has accelerated the R&D of local biotechnology leaders such as Human Genome Sciences, NIH and MedImmune, as well as growing Maryland start-ups such as Martek Biosciences and Digene Corporation. The BSF has various bioreactors from 2 to 250 Liters and well as a range of upstream and downstream process related equipment including; process centrifuges, cell disruptors, tangential flow filtration devices, chromatography systems and freeze driers.	Ben Woodard. Facility Director Biotechnology Research and Education Program 2103 Potomac Bldg. #092 University of Maryland College Park, MD 20742-3371 USA Ph: +1 301.405.3909 Email: [e-mail] Wed: www.biotech.umd.edu/bsf.html
Biologic Therapy Core Facility (BTCF)	The BTCF is an FDA-registered modern cleanroom facility for the manufacture of custom biologics and drugs under GMP conditions for use in clinical trials. The facility provides assistance with IND submissions, manufacturing of both cellular and non-cellular biotherapeutics, and cell selection therapy in a controlled cGMP compliant environment. The BTCF consists of 6 independent (ISO Class 7) manufacturing suites under positive/negative pressure ensuring a safe environment for the production of custom products and rooms can be customized for specialized equipment. The Facility is continuously monitored by an automated system and products monitored in cGMP storage under a variety of conditions.	Dennis Hallahan, MD, facility director William Swaney, MS, operations director Siteman Cancer Center Washington University School of Medicine 660 S. Euclid Avenue Campus Box 8007 St. Louis, MO 63110 USA Ph [phone] or e-mail E-mail: [e-mail] Web: www.siteman.wustl.edu/GMPfacility.aspx
Center for Bioengineering and Biomanufacturing Commercialization	The Center for Bioengineering and Biomanufacturing Commercialization builds an alliance of academia and industry to integrate research, development, commercialization and workforce development.	Dr. Elizabeth Murray Marshall University, Byrd Biotechnology Science Center 241G Huntington, WV 25755 USA Ph: [phone] E-mail: [e-mail] Web: www.science.marshall.edu/murraye/momsite/cbbc.html
Waisman Biomanufacturing.	Waisman Biomanufacturing currently operates and maintains seven cGMP compliant cleanroom areas to accommodate clinical production of mammalian and microbial therapies and aseptic filling of final products. The facility was designed to maximize regulatory compliance and environmental quality and as such provides specific zones or defined areas for each type of activity with appropriate attributes to support and segregate different manufacturing processes. The entire clinical manufacturing facility is key-access controlled and is isolated from other operations within the Waisman Center. The facility has a range of equipment to support the production of recombinant mAbs including; Wheaton Science Model 1862 roller bottle apparatus, single use glass stirred tanks (36 liter capacity), CYTO3 Bioreactor, preparative liquid chromatography systems, scalable single-use tangential flow filtration systems, Sorval RC3BP centrifuge and parallel plate single-use cell culture vessels (multi-tray cell factories) for adherent cell lines	John Keach (Waisman Biomanufacturing Business Development Manager). Waisman Center University of Wisconsin-Madison 1500 Highland Avenue Madison, WI 53705-2280 USA Ph: [phone] Email: [e-mail] Web: www.GMPbio.org
Development Center for Biotechnology, Biopharmaceutical Pilot Plant Facility (DCB)	The Development Center for Biotechnology (DCB) of Taiwan is a nonprofit organization founded in 1984. Co-sponsored by government grants and private donations, DCB's purpose is to advance Taiwan's biotech-pharma industry by building infrastructure, developing key technologies, and training talented workforces in coordination with government, industry, and academic institutions. The facility is cGMP accredited by the FDA and also offers GLP assay development and product characterization capabilities. The facility has 2 × 500 L mammalian cell bioreactors and a suite of corresponding downstream processing equipment.	Dr. Wei-Kuang Chi (cGMP) and Dr. Ta-Tung Yuan (Analytical) 101, Lane 169, Kangning St., Xizhi Dist, New Taipei City 22180, Taiwan, R.O.C Ph: [phone] E-mail: [e-mail] Web: www.dcb.org.tw
National Biologics Facility AIBN Biologics Facility (University of Queensland) Recombinant Protein Production and Purification Facility (CSRIO)	The NBF provides the infrastructure and biotechnology know-how to enable Australian academic and industrial researchers to bridge the gap between laboratory experiments and production of material for preclinical and clinical trials. The NBF is housed in a purpose-built, $13 million facility consisting of laboratories, clean rooms, state-of-the-art equipment and expertise. Staff can advise on and execute all stages of biopharmaceutical preclinical development, from the generation of well-characterized cell lines to the design of bioprocesses suitable for large-scale production and purification. This facility has a range of mammalian cell bioreactors up to 100 L scale and associated downstream processing equipment. The NBF contains a second node located at CSIRO in Melbourne Australia. This group has experience bioprocess optimization.	Dr. David Chin Australian Institute for Bioengineering and Nanotechnology. The University of Queensland. Bldg 75, St. Lucia QLD 4072, Australia. Ph: [phone] E-mail: [e-mail] Web: www.aibn.uq.edu.au/index.html?page=70317&pid=16574 George Lovrecz (CSIRO) Email: [e-mail] Web: www.csiro.au/places/Recombinant-Protein-Facility.html
Centre for Therapeutics Discovery (CTD)	The CTD, based in Mill Hill, translates innovative biology into lead stage therapeutic assets with clear preclinical and clinical developmental pathways. The CTD develops innovative assays and medicinal chemistry programs from high quality hits in collaboration with scientists and clinicians from the MRC and other academic organizations, many of whom are world leaders in their field. The Therapeutic Antibody Group (part of CTD) has substantial antibody engineering know-how, and can deliver humanized antibodies within 6 months. The Therapeutic Antibody Group has substantial antibody engineering experience. It has humanized over 40 antibodies, of which 11 have been progressed to the clinic and 2 (Actemra^® and Tysabri^®) are marketed, and created new therapeutic antibodies.	MRC Technology 1-3 Burtonhole Lane London NW7 1AD United Kingdom Ph: [phone] E-mail [e-mail] Web: www.mrctechnology.org/about/our-structure/centre-for-therapeuticsdiscovery

Note: This table excludes protein expression faculties that such as the Oxford Protein Production Facility UK and other institute based core faculties that are used for production of proteins for internal use, at small-scale or for basic research such as structural studies. The list also does not include the large number of cGMP faculties that have been developed to aid cellular and gene therapy based research and clinical trials. Abbreviations: BSL, biological safety level; cGMP, current good manufacturing practices; CMC, chemistry, manufacturing and controls; DMF, drug master files; FDA, Food and Drug Administration; IND, investigational new drug application; ISO, International Organization for Standardization; NCI, National Cancer Institute; STR, stirred tank reactors; QA, quality assurance; QC, quality control.

Table 3 Analytical characterization of mAbs

Product characteristic	Property	Commonly used Analytical Techniques
Physical and chemical characteristics	Purity	Electrophoresis, Reverse phase HPLC, Size exclusion HPLC
	Integrity/size	Electrophoresis, Mass spectrometry, size exclusion HPLC, dynamic light scattering
	Identity	Isoelectric focusing, Peptide mapping, Ion exchange HPLC, Mass spectrometry
Potency/activity	Antigen binding	Immunoassay (ELISA), Biacore (SPR), Micro-calorimetry (ITC and DSC)
	Bioassays	Cell proliferation, Complement mediated cytotoxicity, Reporter-gene assays
Product related impurities	Product related variants e.g., aggregates, glycosylation, amino acid modifications	Electrophoresis, Size exclusion HPLC, Mass spectrometry
	Product related substances, e.g., breakdown products	Electrophoresis, Reverse phase HPLC
	Host Cell Protein	Immunoassay
	Host Cell DNA	DNA hybridization, qPCR, DNA binding-threshold fluorescence (e.g., picogreen)
Process related impurities and contaminants	Leeched Protein A	Immunoassay (ELISA)
	Cell culture medium, feeds and supplements	Immunoassay (ELISA), Reverse phase HPLC
	Viruses	qPCR, Electron microscopy, In vivo and in vitro assays
	Microorganisms	Bioburden (plate and broth based), Endotoxin—LAL test
	Others: column/vessel leachates/extractables cell culture medium components, purification reagents/chemicals	Reverse phase HPLC, ion exchange chromatography, GC-MS, NMR

Physical characterization of recombinant mAbs is crucial at early stages in development (Adapted from Flatman et al.66).

Flatman S, Alam I, Gerard J, Mussa N. Process analytics for purification of monoclonal antibodies. J Chromatogr B Analyt Technol Biomed Life Sci 2007; 848:79 - 87

 PubMed Web of Science ®Google Scholar