SBIR-STTR Award

In today's biopharmaceutical pipeline, monoclonal antibodies (mAbs) are a predominant modality for a broad range of clinical indications including oncology and inflammatory diseases. Most antibody therapies have to be administered to the patients in large doses of 4-15 mg/kg, resulting in extremely high cost for patients (~$2,000-$5,000 per dose; up to $100,000 per year). Therefore, manufacturing capacity and cost per purification campaign are two critical factors for making these therapies more affordable. In recent years, new developments in protein expression, cell culture, and fermentation technologies have led to significant advancements in protein production and antibody titers. The development of the subsequent downstream purification steps, however, has not kept a similar pace and can become a bottleneck that impedes achieving a cost-effective and robust manufacturing process. Currently, the downstream processing accounts for up to 70% of mAb production cost. As such, scalable technologies that create manufacturing equipment with in- creased capacity through downstream process productivity improvements and cost reduction are highly de- sired. In mAb purification, the "industry-standard" Protein A capture step is the most expensive step, contributing 51% to the overall purification cost [and up to 40% of the total cost per grammAb]. We propose to demonstrate the purification of a mAb using a highly efficient Protein A capture step with a novel continuous chromatography process and instrument and to demonstrate up to 10-fold savings in Protein A chromatography media and buffers as compared to the traditional single-column batch method. Our novel chromatography instrument is a bench top multicolumn system capable of simulated moving bed (SMB) chromatography proto- cols that increase productivity up to 10-fold vs. conventional single-column methods. In Phase I, we will purify a model mAb using our instrument and the Step-SMB Protein A capture process. We will compare the efficacy of this process in terms of yield and purity to that of the traditional single-column process purified mAb. We will also identify instrument hardware and software design changes to insure the optimized SMB process would achieve FDA-recommended purity specifications. In Phase II, we will extend this demonstration to a prototype manufacturing process supporting up to eight-step cGMP Protein A capture step of mAb, implement in-process controls, such as UV absorbance, pH and conductivity, and evaluate the effect of process throughput on mAb purity. We will also work with a commercial partner to integrate our continuous mAb Protein A capture step into a complete purification process.

Public Health Relevance:
In recent years, new developments in protein expression, cell culture, and fermentation technologies have led to significant advancements in protein production and antibody titers. The development of the subsequent downstream purification steps has not kept a similar pace and is becoming a bottleneck that impedes achieving a cost-effective and robust manufacturing process. Currently, the downstream processing accounts for a significant percentage of mAb production cost, and improvements in manufacturing technologies which increase capacity and productivity are highly desired for speeding up progress through trials and, ultimately, improving patient health and access to lower cost therapeutic mAbs. Protein A capture is the "industry-standard", most efficient yet expensive step in mAb purification, and increasing productivity while reducing cost of this step would lead to more affordable biological drugs for patients.

Public Health Relevance Statement:
In recent years, new developments in protein expression, cell culture, and fermentation technologies have led to significant advancements in protein production and antibody titers. The development of the subsequent downstream purification steps has not kept a similar pace and is becoming a bottleneck that impedes achieving a cost-effective and robust manufacturing process. Currently, the downstream processing accounts for a significant percentage of mAb production cost, and improvements in manufacturing technologies which increase capacity and productivity are highly desired for speeding up progress through trials and, ultimately, improving patient health and access to lower cost therapeutic mAbs. Protein A capture is the "industry-standard", most efficient yet expensive step in mAb purification, and increasing productivity while reducing cost of this step would lead to more affordable biological drugs for patients.

NIH Spending Category:
Bioengineering; Biotechnology; Breast Cancer; Cancer; Immunization; Orphan Drug; Rare Diseases

Project Terms:
Accounting; Affinity; Affinity Chromatography; alemtuzumab; Antibodies; Antibody Formation; Antibody Therapy; Avastin; Beds; bevacizumab; Binding (Molecular Function); Biological; Biological Products; Breast Cancer Treatment; Buffers; cancer therapy; cancer type; Cell Culture Techniques; Cells; Cetuximab; Chromatography; Chronic Lymphocytic Leukemia; Clinical; Clinical Trials; Colorectal; Colorectal Cancer; comparative efficacy; cost; cost effective; Cost Savings; Cyclic GMP; design; Development; Devices; Disease; Dose; drug production; Equipment; Erbitux; feeding; Fermentation; Glioblastoma; Guidelines; Head and Neck Cancer; Health; Healthcare Industry; Human; Immunotherapy; improved; in vitro activity; Industry; Inflammatory; innovation; instrument; Lead; Legal patent; Life; Link; MabCampath; malignant breast neoplasm; manufacturing process; Marketing; Measures; meetings; Methods; Modality; Modeling; Monoclonal Antibodies; Monoclonal Antibody Therapy; Non-Hodgkin's Lymphoma; Non-Small-Cell Lung Carcinoma; novel; Nucleic Acids; oncology; Patients; Pharmaceutical Preparations; Phase; Plant Resins; Price; Process; Production; Productivity; protein expression; Proteins; Protocols documentation; prototype; Renal carcinoma; Research Personnel; rituximab; Roche brand of rituximab; Roche brand of trastuzumab; Sampling; Savings; Simulate; Software Design; software systems; Speed (motion); System; Technology; Therapeutic Monoclonal Antibodies; Time; Trastuzumab; Vaccines; Work

Monoclonal antibodies (mAbs) have become the leading drug class for cancer treatment, and over 150 anti-neoplastic mAbs are in the biopharmaceutical pipeline. Most mAb therapies must be administered in doses of 4-15 mg/kg, resulting in per patient costs of up to $100,000 per year. The high price of mAb therapy causes a significant financial burden for patients, insurance companies and the health care system. To be- come practical for the health care industry, the cost of developing and producing these drugs must be reduced. Clinical manufacturing expenses directly contribute to the overall drug development costs. Recent developments in protein expression, cell culture, and bioreactor technologies have led to substantial advancements in protein production and mAb titers. The development of downstream purification processes has not kept pace and represents a bottleneck that impedes cost-effective and robust manufacturing. Currently, downstream processing (DSP) accounts for up to 70% of the total mAb production cost. The "industry-standard" Protein A capture (PAC) is the most expensive DSP step, contributing up to 40% of the total cost per gram of product. The current PAC method still uses one large column in a sequential batch process. Continuous SMB (simulated moving bed) chromatography offers significant advantages over standard batch methods, including more efficient use of expensive adsorbent with smaller columns, reduced buffer consumption, and operation under steady-state conditions, allowing more robust process analytics. In the Phase I SBIR project (R43 CA162632-01) we investigated the feasibility of developing a continuous PAC process for the purification of clinical-grade mAbs using our lab-scale Octave(tm) SMB System. We obtained equivalent or better mAb purity when our continuous process was directly compared with the standard batch process. How- ever, the current Octave System is not designed for cGMP compliance or the scale required for clinical manufacture. In Phase II we propose to develop a large-scale continuous chromatography device for economical purification of clinical-grade antibodies, based on the Octave valve design and our Phase I findings. The Phase II device will support flow rates up to 2 L/min and processing of 500 L culture fluid containing 5-10 g/L mAb in 8-20 hours, which matches the future demand for therapeutic mAbs in the range of 200 kg/yr. The Phase II de- vice and optimized PAC process will increase productivity at least 3-fold relative to batch methods, and will feature a single-use flow path to eliminate the need for sanitation and revalidation between campaigns. This Phase II project will develop at least one prototype device that will be placed at a Beta test site. The device will be a customizable plug-and-play chromatography module compatible with future integrated continuous bioprocessing facilities. This project fits with the 2011 FDA strategic plan, which seeks development of improved product manufacturing technologies including continuous processes rather than batch approaches.

Public Health Relevance Statement:


Public Health Relevance:
Recent developments in protein expression, cell culture, and bioreactor technologies have led to substantial improvements in protein production and increased antibody (mAb) titers. The development of downstream purification processes has not kept pace and represents a bottleneck that impedes cost-effective and robust manufacturing. Currently, downstream processing accounts for a significant percentage of the total mAb production cost. Improvements in manufacturing technologies which increase capacity and productivity are highly desired for speeding up progress through clinical trials. The device developed under this proposal will dramatically improve productivity and reduce cost of the most expensive purification step by up to 80%. Incorporation of this device into manufacturing processes will ultimately improve patient access to new anti-cancer mAb therapies at lower cost.

Project Terms:
Accounting; Achievement; Antibodies; Automation; base; Beds; Biological Products; Biomanufacturing; bioprocess; Bioreactors; Boat; Buffers; cancer therapy; Cell Culture Techniques; Chemicals; Chromatography; Clinical; Clinical Trials; Computer software; Consumption; cost; cost effective; Cyclic GMP; design; Development; Devices; Dose; drug development; Electronics; Environment; fluid flow; Future; Guidelines; Healthcare Industry; Healthcare Systems; Hour; Housing; improved; Industry; Insurance; Insurance Carriers; Ion Exchange; Legal patent; Liquid substance; Logic; Malignant Neoplasms; Manufacturer Name; manufacturing process; meetings; Membrane; membrane assembly; Methods; Monoclonal Antibodies; neoplastic; operation; Patients; Pharmaceutical Preparations; Phase; Play; Price; Process; Production; Productivity; protein expression; Proteins; Protocols documentation; prototype; public health relevance; Pump; Relative (related person); Research; Sanitation; seal; sensor; Simulate; Site; Small Business Innovation Research Grant; software systems; Speed (motion); Sterile coverings; Sterility; Strategic Planning; Stream; System; Technology; Test Result; Testing; Therapeutic Monoclonal Antibodies; Voice