Background on Live Biotherapeutics
The community of microorganisms that reside in various regions on and within the human body is known as the microbiome. These microbes produce and consume numerous compounds that considerably impact our health and well-being. There is a delicate balance in our coexistence with these microorganisms, and when the community balance is perturbed, it can lead to various health complications, such as recurrent Clostridium difficile infections in the gut. The reintroduction of beneficial microorganisms has the potential to act as clinically relevant therapies to re-establish and maintain a healthy microbiome. Large consortia of bacteria, as well as individual bacterial strains, are currently being studied as potential agents for treating disease. Some of the disease states currently targeted by Live Biotherapeutic Products (LBPs) include gut, dermatologic, neurologic, immunologic, reproductive, and oncologic disorders. Some of the strains we have worked with may be reviewed by clicking on the icon below.
List Labs is flexible to meet the unique needs of LBP client projects. We offer the following development services to establish a reproducible and robust process:
- Cell Bank Preservation
- Inoculum/Seed Optimization
- Media Optimization
- Fermentation Development
- Harvest Development
- Lyophilization Development
- Analytical Method Development
- Process Scale-Up
Initially, a technical transfer is desired to acquire familiarity with the specific bacterial strain and replicate the customer’s data and results. Small scale work is designed with the goal of a seamless transition toward a process that will support the large-scale manufacturing process, compliant to cGMP. Optimization of media and fermentation conditions frequently guarantee an increased yield of the product. In addition, animal-derived components are replaced with non-animal origin (NAO) components in preparation for cGMP manufacturing. Various fermentation process parameters can be evaluated in parallel in a scalable model to establish a proper design space for the manufacturing process including:
- Media Formulations
- pH Control
- DO Control
- Agitation Speed
- Gas Sparging
- Incubation Temperature
- Fed-batch Parameters
To support process development and future process validation, clients are recommended to provide in-process tests, to aid in the evaluation of each step’s success. Test methods for a variety of assays are currently in use, from Colony Forming Unit (CFU) determinations to measuring the bioactivity of the organism or the compounds it produces. Harvest development entails optimization of the hollow fiber membrane surface area, flow rate, and pore size to concentrate the bacterial cells and remove fermentation byproducts. This process presents a significant challenge for strict anaerobes which are expected to be viable after processing. List Labs has developed proprietary methods with anaerobes to recover cells while maintaining an anoxic environment to ensure minimal loss.
In order to preserve the harvested bacteria, lyophilization, or freeze-drying, is typically preferred. List Labs has used this process to successfully preserve bacterial cultures for more than forty years. When bacteria are used as therapeutics, maintaining a high density of viable organisms
that are stable over a reasonable product lifetime is desired. For lyophilization, the concentrated bacteria are suspended in a compatible solution that will include a cGMP compliant cryoprotectant. The lyophilization process involves a series of well-designed steps for flash freezing and the controlled application of heat while under vacuum to produce a dry, stable product with high viability. Several formulations can be tested simultaneously, allowing for rapid process development. At List Labs, we have the experience to design these processes and the analytical tools to evaluate cycle success.
Analytical method development is vital to the success of your project. Projects will require analyses specific to the desired product as well as generally applicable analyses described in the USP. These tests may include moisture, water activity, pH, osmolality, endotoxin content, viability, bioburden, and bioactivity. We can apply our considerable experience with suitability testing to develop the necessary analytics specific to your project.
Our highly skilled team has extensive experience with cGMP manufacturing of LBP products from a broad range of genera, including strict anaerobe species. Our expertly designed manufacturing facility provides containment in ISO7 or ISO8 suites. In addition, we have multiple suites allowing parallel cGMP manufacturing activities, streamlining project timelines, and avoiding delays.
List Labs offers state of the art cultivation of strict anaerobes for Research Cell Bank or Master Cell Bank production in an internal HEPA filtered anaerobe chamber to maintain an aseptic process not possible in typical anaerobe chambers. List Labs has several large-scale options for cultivation and fermentation of your organism including a 100L stainless steel vessel with validated cleaning procedures and 40L disposal bioreactors. A closed system is utilized to harvest the microorganism by tangential flow filtration using a hollow fiber prior to formulation and lyophilization. Our production capabilities include three lyophilizers with a 5 tray, or 3000 x 2mL vials, capacity each. In addition, we have experience with sieving and powder filling into different packaging modalities and applicators. We offer the following manufacturing services:
- Research Cell Bank Production
- cGMP Master and Working Cell Bank Production
- cGMP Drug Substance Production
- cGMP Drug Product Production
- Fermentation and Harvest Services
- Formulation and Lyophilization Services
- Sieving and Dry Filling Services
- Liquid Filling Services
- Custom Packaging Services
List Labs staff has over 25 years’ experience manufacturing bacterial based novel drug substances and products. List maintains several Drug Master Files with FDA and Health Canada and provides support to clients for IND filings. As required by the State of California, List Labs maintains a current Drug Manufacturing License and CDC Select Agent registration enabling us to produce drug substance and drug products for our clients.
List Labs has manufactured a variety of bacterial toxins and proteins, live biotherapeutics products (LBS), and polysaccharides for our clients. cGMP manufacturing has included both drug substance and drug products for Phase I and II clinical trials.
List complies with 21 CFR 210, 211, as well as ICH and Regulatory Guidelines, as applicable, for early-stage Investigational New Drugs (IND) and Investigational Medicinal Products (IMP).
For more information, see our GMP services page.
QC Analytical Testing
The majority of QC testing for LBPs can be developed and executed by List Labs, creating a seamless full-service offering for your project requirements. Typically testing provided by List Labs for Live Biotherapeutic Products includes:
- Colony Forming Units (CFU)
- Live Dead Fluorescence Assays
- Total Cell Counts
- Bioactivity Assays
- Moisture Analysis
- Water Activity
- Endotoxin Content
- Colony Morphology
- Cell Morphology
- Gram Stain
- Strain Identification
- Bioburden (USP<61> and USP<62>)
For more information about our QC analytical testing, click the icon below.
Equipment in our facility for manufacturing LBPs includes anaerobe chambers, incubators, stainless steel fermenters, disposable bioreactors, tangential flow filtrations systems, lyophilizers, dry chambers, and fill equipment. For more information about our available equipment, click on the icon below to view our full equipment list.