Microbial Cell Line Development

Raising the Bar on Microbial Expression and Manufacturing for New Biologics

KBI Uses An Early Optimization Workflow To Ensure Manufacturing Success. As A Result, More Molecules Are Manufactured Using An E. Coli Expression System Than Ever Before.

Ahead of upstream process development, KBI focuses on microfermentation screening for microbial cell line development (CLD) to yield consistent, high titers across strains and molecules. At KBI, our years of vertical process integration knowledge and data can allow us to scale directly into 10L stainless steel or 30L single-use systems - ahead of further scale-up.

We quickly develop robust, scalable protein manufacturing processes through in silico protein analysis, combined with wet lab IP and industry-leading screening workflows. Our microbial CLD services include:

CLD with E. coli expression systems, leveraging our PUREplatform™
Protein refolding and scalable processes for difficult-to-refold products
High-density fermentation with rapid biomass production cycle times

Introducing KBI's PUREplatform™, featuring PUREcoli™

A pure microbial strain for clean, efficient therapeutic protein production. As part of the KBI PUREplatform, PUREcoli is expertly paired with PUREplasmids™ and PUREmedia™ and our cutting-edge fermentation process that fits together into a complete microbial CLD service package, catered to your molecule and business needs.

Learn More about PUREcoli >>

Biologics manufacturing by high-density microbial fermentation has advantages of development speed, low cost, and scalability.

Molecule manufacturability assessment and microbial optimization are the start of KBI's approach to advanced microbial cell line development. These steps sets your project up for success by leveraging the known advantages of microbial fermentation.

KBI's approach is the best fit when selecting E. coli as the preferred host for expression of non-glycosylated proteins, including both soluble and insoluble product accumulation. This includes downstream capture, host-cell impurity clearance, and where necessary, controlled and scalable refolding. KBI also has experience with various non-E. coli bacterial hosts such as Pseudomonas-based and third-party proprietary expression systems.

In cases of insoluble protein accumulation in inclusion bodies (IBs), KBI develops high-yielding recovery processes with analytical characterization of conversion to an active product by solubilization and refolding techniques. KBI's unique expertise in this area has enabled the use of microbial hosts for the expression of several known difficult-to-refold products at a favorable scale and cost.

KBI applies proprietary proven, well-characterized microbial recombinant protein expression systems to provide state-of-the-art, early-stage microbial cell line development through cGMP services, including:

Proprietary PUREcoli strain evaluations include:

Arabinose, rhamnose, phosphate depletion, and IPTG-inducible gene promoters
Removable leader sequences for localizing proteins, increasing expression, or editing the N-terminus
Soluble protein accumulation
Insoluble protein/inclusion bodies (IBs) accumulation
Periplasmic protein accumulation
Gene and codon optimization of the expression plasmids
High throughput cell line selection based on fermentation performance, product yield, and product quality
Options to integrate early analytical characterization into the clone selection process, e.g., state-of-the-art mass spectrometry
Understanding of business and science-driven objectives related to the expression platform choice, e.g., product quality, IP status, and licensing terms
Implementing ICH Q5D and ICH Q5B guidelines

Proof-of-concept (POC) screening and early-stage material generation

Preclinical programs designed to meet your molecule needs business expectations
Cell line development services are offered as vertically integrated programs for higher speed and cost-efficiency from CLD through to cGMP manufacturing and fill finish, but can also be performed as stand-alone services.
Selected cell line(s) generate Research Cell Banks (RCBs) that are the foundation for further development and scale-up.

Full-scope process development

Master Cell Bank (MCB) generation, testing, and characterization are outsourced to established, qualified, and quality-audited third-party vendors.
High-throughput high-density fermentation optimization
Scalable harvest and recovery process optimization
Downstream chromatography screening and optimization of orthogonal purification strategies
Pilot-scale confirmation and GLP-enabling material generation

cGMP manufacturing, PPQ, and Commercialization

300L single-use fermentation and 2000L stainless steel fermentation scale options
Existing open capacity with US-based expansion planned for near future
PPQ expertise to prepare your product for a successful commercial launch
Facility with existing commercially licensed products with favorable inspection outcomes
Fully staffed quality and regulatory departments providing inspection-ready status at any time
Access to real-time data trends and highly qualified Manufacturing Sciences staff to ensure the process runs smoothly while in the plant
May provide a PiP (person-in-plant) during at-scale production

Microbial Cell Line Development