Cell Line Development

Cell line development is the process by which the cellular machinery is co-opted to manufacture therapeutic biologics or other proteins of interest. You can use different expression systems for cell line development: bacterial, plant-based, yeast, mammalian. Most commonly used for the production of the complex protein is Chinese hamster ovary (CHO), cells grown in suspension cultures for eventual use in bioreactors at the manufacturing stage.

Cell Line Development Process

The development of these cell lines is a long and arduous process with numerous steps and challenges (Figure 1). Once CHO cells have been transfected with the construct of interest a selection step is required to enrich for those cells that have taken up the construct. Single cells must then be isolated, grown into colonies, and assayed for protein production. Once this production screen has been accomplished, the top candidates are expanded and maintained to test for expression stability. Eventually the top hits are further optimized in different media conditions to enhance expression or other protein attributes (e.g. glycosylation) before scaling up to production levels. Because this process takes months to complete, there are many more challenges than a typical short-term cellular workflow. First, maintaining sterility is essential during various manipulations and throughout the weeks of growth from a single cell. Secondly, as with most screens, the throughput required to identify a handful of optimal clones typically requires thousands of wells. This throughput then tapers off after the hit picking phase, so one should schedule overlapping campaigns to maximize resources. Finally, as clones move through the system and are reformatted into lower density plates during hit picking, all previously recorded data must be tracked as these cell lines are established. We have overcome these challenges by automating the establishment and screening of clones for cell line development.

Products for Cell Line Development

Biomek iSeries

Automate every step of the complex Cell Line Development workflow - from transfection to enrichment to limiting dilution to culture/feeding to assaying to hit-picking and expansion.

MoFlo Astrios

Provides an automated, high throughput single cell depositing system. This process is mandatory for master cell bank generation.

Monochronality verification system (Isolate single cells and expand)
Validate single cell per well

Vi-CELL BLU

Used at multiple steps along cell line development. Eliminate human error and user bias when counting cells and determining viability.

Cell count is critical to determine cell expansion and passaging
Cell viability- kiow how many cells are live and how many are dead

Avanti J-15R

Separate cells from culture media for antibody titer assays, quantification.

Separate cells from media for antibody titer
Pellet cells for media exchanges

Application Resources

Title	Description
Automating Cell Line Development {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Cell line development is the process by which the cellular machinery is co-opted to manufacture therapeutic biologics or other proteins of interest. This most frequently involves the production of antibodies in Chinese hamster ovary (CHO) cells grown in suspension cultures for eventual use in bioreactors at the manufacturing stage. The development of these cell lines is a long and arduous process with numerous steps and challenges.
Mastering Cell Counting {C00ACFAF-8661-4DF6-8BA5-A7A099A7BA9C}	Cell line development is a highly-involved, multi-week workflow that involves the plating, screening, and expansion of single cell-derived clones to establish a protein-expression cell line. This complex process required significant throughput of sterile cell manipulations, long-term incubations, centrifugation, cell counting, monoclonality determination and growth tracking, and protein production assessment.
Cellular Challenges: Taking an Aim at Cancer {C00ACFAF-8661-4DF6-8BA5-A7A099A7BA9C}	Immunotherapy is a powerful treatment that harnesses the body’s immune system in the fight against cancer. With optimized cell therapy, the result is an expanded population of T cells primed to recognize and eradicate malignant tumor cells that would otherwise escape immune detection.
Automating Cell Line Development for Biologics {C00ACFAF-8661-4DF6-8BA5-A7A099A7BA9C}	Cell line development is a highly-involved, multi-week workflow that involves the plating, screening, and expansion of single cell-derived clones to establish a protein-expression cell line. This complex process required significant throughput of sterile cell manipulations, long-term incubations, centrifugation, cell counting, monoclonality determination and growth tracking, and protein production assessment.
Cell Line Development {9E29D7A5-B5EC-470A-A5C1-4EF9BFAE9959}	Traditionally, cell line development has been achieved manually, limiting it to low-throughput applications and making it susceptible to variability caused by human error and contamination. Automating the cell line development workflow— from a point of genetic manipulation through clone selection—increases throughput and optimizes media conditions which are critical for cell lines needed for vaccines, antibody production, recombinant therapies and drug screening.
Method for Determining Cell Type Parameter Adjustment to Match Legacy Vi CELL XR {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note details how you can use the plate loader feature on the Vi-CELL BLU to match cell types to give cell counts and viability levels equivalent to those obtained on existing Vi-CELL XR instruments.
Considerations of Cell Counting Analysis when using Different Types of Cells {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This paper reviews several different cell types and cell preparation approaches using standard cells, such as, CHO cells, EL4 cells, SF9 cells, and HELA cell cultures.
Evaluation of Instrument to Instrument Performance of the Vi-CELL BLU Cell Viability Analyzer {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note details one of the key strengths of the Vi–CELL BLU, the ability to fine tune the performance to minimize variability between different instruments. This is particularly valuable in facilities that utilize multiple instruments but can also be important when instruments are occasionally shared between departments.
Cell Line Development – Limiting Dilution {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Limiting dilution is a traditional approach to achieve monoclonality. It entails diluting cells to the level that there is one cell on average per unit of volume plated in the wells. However, as predicted by the Poisson distribution, roughly one third of the plated wells will have zero cells, one third of wells will have a single cell, and one third will have multiple cells per well (Coller and Coller 1986).
Cell Line Development – Hit Picking {442DF88C-CA7F-4B78-AF75-9D2B985534C0}
Cell Line Development – Selection and Enrichment {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Cell line development is a highly involved and inefficient process. The quest to find cells that stably express and secrete antibodies or other protein products from a transfected pool of cells requires the screening of thousands of wells/cells.
Cell Line Development – Data Handling {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Cell line development is one of many workflows that has significant data challenges. Like any screening process, the sample throughput provides a challenge from the sheer amount of data. This becomes even more challenging when multiple analytical instruments are used to generate different types of data, often in different formats.
Efficient Factorial Optimization of Transfection Conditions {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Nucleic acid transfection is a procedure used in nearly any cellular laboratory and the abundance of commercially available reagents has made this a seemingly simple endeavor. However, whether one is transfecting plasmid DNA or inhibitory RNAs, different cell lines can have significant differences in the ease of transfection or cellular survival. Determining the optimal plating conditions for high transfection efficiency and low cytotoxicity can be highly involved.
Cell-Line Engineering {C00ACFAF-8661-4DF6-8BA5-A7A099A7BA9C}	Bioproduction is a booming industry, with more therapeutics being produced in cell culture than ever before. With increased workflow flexibility, decreased variability, and high product yields, cell-line engineering is here to stay. But before you take your bioproduction to the next level, you’ll need to know a bit more about your product, your cells, and their health. It’s time for a cell-line engineering check-up
Cell Line Development {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	As biologics research continues to evolve and gain prominence in therapy development for disease states, the need for reliable and robust cell lines will greatly increase
Automating Cell Line Development {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note features automating cell line development with a Biomek i7 liquid handling workstation and integrated devices in order to overcome challenges for this workflow
Cell Line Development – Limiting Dilution {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	In this application note we describe the automation of limiting dilution on a Biomek i7 liquid handling workstation that overcomes a number of challenges associated with this workflow
Evaluation of Instrument to Instrument Performance of the Vi-CELL BLU Cell Viability Analyzer {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note examines the comparability of results from multiple Vi-CELL BLU cell viability analyzers.
Cell Line Development – Limiting Dilution {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Limiting dilution is a traditional approach to achieve monoclonality
Automated Solutions for Cell Line Development {A7DF4F56-8AF9-4BF1-88BE-766AB73FBC6B}	Your Guide to Workflow Intelligence
Automating Cell Line Development for Biologics {C00ACFAF-8661-4DF6-8BA5-A7A099A7BA9C}	Cell line development is a highly-involved, multi-week workflow that involves the plating, screening, and expansion of single cell-derived clones to establish a protein-expression cell line
Method for Determining Cell Type Parameter Adjustment to Match Legacy Vi-CELL XR {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note assists in matching Vi-CELL BLU cell types to give cell counts and viability levels equivalent to those obtained on existing Vi-CELL XR instruments.
Instrument-to-Instrument Variability in the Vi-CELL Automated Viability Analyzer {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Authors: Clarence Lew, Senior Applications Scientist, John A. Gomez, Senior Staff Technical Operations Engineer, Matthew N. Rhyner, Ph.D., Global Marketing Manager-Analytical Products \| Beckman Coulter Life Sciences, Brea, CA USA \| Published 2012
Considerations of Cell Counting Analysis when using Different Types of Cells {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	This application note reviews several different cell types and cell preparation approaches using standard cells.
Choosing the Right Instrument {9E29D7A5-B5EC-470A-A5C1-4EF9BFAE9959}	for Cell Counting and Particle Characterization
Cell Line Development {9E29D7A5-B5EC-470A-A5C1-4EF9BFAE9959}	Automating the cell line development workflow increases throughput and optimizes media conditions which are critical for cell lines needed for vaccines, antibody production, recombinant therapies and drug screening
Cell Line Development – Hit Picking {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	The basic concept of hit picking is simple – 1) identify hits based on assay data and 2) consolidate these hits into a new labware in an identifiable way
Efficient Factorial Optimization of Transfection Conditions {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Nucleic acid transfection is a procedure used in nearly any cellular laboratory and the abundance of commercially available reagents has made this a seemingly simple endeavor
Cell Line Development – Selection and Enrichment {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	The quest to find cells that stably express and secrete antibodies or other protein products from a transfected pool of cells requires the screening of thousands of wells/cells
Automating Cell Line Development {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Cell line development is the process by which the cellular machinery is co-opted to manufacture therapeutic biologics or other proteins of interest
Cell Line Development – Data Handling {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	Cell line development is one of many workflows that has significant data challenges
Cell Line Development – Hit Picking {442DF88C-CA7F-4B78-AF75-9D2B985534C0}	In this application note we discuss how hit picking was automated as part of a cell line development workflow
Vi-CELL® XR Cell Type Tutorial {B14306BD-BB46-4B27-BF3E-B11862085BC7}	Introduce users to the Cell Type Feature in the Vi-CELL XR software