Biologics are complex molecules which must be manufactured in living cells. "Host cells" are genetically edited to produce (secrete) a desired protein/antibody, which in turn is harvested, purified (= biologic) and then used to treat a patient. Genetically editing host cells involves opening their cell membrane to allow the gene to enter.
Most drugs you use today are known as small compounds, e.g. Aspirin. They are tiny molecules that can activate or inhibit a cellular function and can be synthesised in a chemistry lab. But in the last 40 years, science has learned to harness this new type of drug - the biologic. Biologics are much larger compounds, that have exponentially more possibilities in the body.
As well as affecting cellular pathways, they can actually carry out biological functions themselves. This means they can influence diseases where an individual may not have the cellular components for an essential function. This is something small molecules will never be able to do. Biologics also do this with a much higher degree of specificity and a significantly lower chance of toxic side effects that small molecules.
The first big biologic on the market was insulin and it has completely changed how the world deals with diabetes. As research continues, more biologic drugs will be produced and there is no doubt that biologics will play a significant role in the future of medicine.
The importance of cell counting for biologics manufacturing
A reliable cell count is crucial for many regulatory aspects of biologics manufacturing:
Safety, efficacy, purity, potency, identity, quality
Oversight of both product and process
Quality control of source materials, intermediates, and product
Reproducibility of lots
Comparability after manufacturing change
HOW DOES IT WORK?
Biologics are difficult to manufacture and since the starting material are cells; the importance of quality control with respect to cell counting, viability and transfection efficiencies cannot be overstated.
One of the biggest bottlenecks within cell manufacturing is Cell Line Development (CLD): the process of turning a regular cell ("host cell") into a highly efficient cell producing the desired protein ("cell line"). To to this, the host cell is opened ("open-membrane") in the presence of the gene; the gene enters the cell and the cell closes as it's membrane seals. The gene genetically engineers or reprograms the host cell and the result is a Cell Line which will produced the desired protein (and thus biologic) stably and with a high yield. Cell Lines are unique to each manufacturer and are the source of all future drug product (Master Cell Bank). They are grown in large bioreactors.
Cell Line Development
The process of CLD, and in particular transfection, can be quite harsh causing significant cell death. Therefore, it is important to analyse the cell population post-transfection for overall cell count and viability.
Cellix's Inish Analyser is the first product on the market that can instantaneously determine if the cell membrane has been successfully opened and thus, whether the gene has been delivered. This provides un-paralleled insight into the transfection process, saving researchers days in unnecessary analysis of dead or non-transfected cells.
WHAT DO I NEED TO GET STARTED?
In the examples below, we highlight results from CHO cells and yeast cells but as genetic editing techniques becomes more widespread, so too are the cells being investigated for biologics production.
Chinese Hamster Ovary (CHO) cells are widely known as the gold standard for the production of biologics, mainly because they have been widely studied in genetics and they grow well in culture. Yeast cells also grow readily in culture and readily accept genetic modifications making them another popular choice for biologic production.
Our Inish Analyser does not require any expensive consumables such as slides or cartridges. Instead we use standard eppendorf tubes. Sample preparation is very simple - just using a pipette.
In general, you will need the following to execute cell counting, viability and transfection efficiency assays:
Inish Analyser: The Inish Analyser provides a simple readout for Cell Counting & Viability assay or Transfection Efficiency Assay. Results are displayed in a table or fcs file format and both are easily exported to a USB or other connected device.
Inish Analyser Buffer: 40µL of cell sample is diluted in 360µL buffer.
Sample and waste tubes, 1.5mL size: we use blue ones for samples and purple ones for waste - this helps avoid making mistakes!
Pipettes & pipette tips:
P-200 yellow pipette (20-200µL) with corresponding yellow pipette tips for sample pipetting.
P-1000 blue pipette (200-1000 µL) with corresponding blue pipette tips for buffer pipetting.
Vortex mixer (optional): not a requirement as you can pipette your sample vigorously instead.
Materials required for Cell Counting & Viability Assay with the Inish Analyser
3 simple steps to get your cell count and viability analysis:
Pipette 40µL of sample into the sample tube.
Pipette 360µL of Inish Analyser Buffer into the sample tube. This provides a 1:10 dilution and gives a total volume of 400µL in the sample tube. Mix by manually pipetting vigorously or use a vortex for a few seconds.
Select the "Cell Count & Viability Assay" on the Inish Analyser touchscreen and follow the instructions.
2 sets of results:
Cell Viability with CHO cells
Cell Viability with yeast cells.