How to Get Reliable Cell Counting and Transfection Analysis for Biological Products?
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How to Get Reliable Cell Counting and Transfection Analysis for Biological Products?


Introduction

The biologics industry is booming so much that the global market will reach USD$749.62 Billion by 2028. If you're part of this growing field, it's worth staying updated about the latest technological innovations so that you can get the most out of your experiments.


The Biologics Heated Market

The use of biologics is skyrocketing in both academic and industry scenarios. This is because scientists are more aware of how useful cellular and gene-based products are in treating human diseases like cancer and rare genetic conditions.

So, the pharma industry is investing heavily in the R&D of biologics on a larger scale. As a result, we have several biologic-based therapies being approved.


A New Perspective on Something Old


Biological products are made from living organisms or parts of them. Although the concept sounds futuristic, humans have been using biological products for many years [1].

With the increased knowledge about diseases, new biological targets appeared, paving the way to develop new therapies. We now have a wide range of biological products available such as vaccines, recombinant therapeutic proteins, gene therapies, natural protein sutures, coagulant factors, and diagnostic tools [1].

Biologics manufacturing usually involves [2]:

  • Cell line construction

  • Upstream processing (fermentation)

  • Downstream purification

  • Drug formulation

Common Cell Types Used in Biologics Manufacturing

Common host organisms for producing biopharmaceuticals include Escherichia coli, mammalian cells, yeasts, and plants. Each one has its advantages and disadvantages depending on the application [3].


Yeast Cells


Yeast cells are a popular choice for cell product manufacturing and gene editing. Engineered yeast is a well-known platform for the synthesis of highly valuable medical compounds [3].

In industrial applications, the most commonly used species are Saccharomyces cerevisiae, Pichia pastoris, Yarrowia lipolytica, and Kluyveromyces marxianus [3].


The main advantages of using yeast cells in biopharmaceutical production are [3]:

  • Fast growth

  • Inexpensive growth media requirements

  • Many species are generally regarded as safe

  • Easy genetic manipulation

  • Compatibility with high-density large-scale fermentation processes for biochemical production.

Mammalian Cells

Mammalian cells like Chinese Hamster Ovary (CHO) cell lines are preferred for post-translational modification. These cells display high yields, but their doubling time is slow. They also require complex media and may be subjected to viral contamination [3].


Other cell types

E. coli grows fast but may form endotoxins and inclusion bodies. Finally, plant cells grow slower and are more difficult to manipulate [3].


Biologic Manufacturing Challenges

The living cells that produce biologics are sensitive and require extra care in fermentation, aseptic processing, storage, and testing [1].

While chemical drugs are usually small molecules, biologics are often complex and heterogeneous compounds. These products are also more prone to microbiological contamination [1].


Master Cell Banks

Another particularity of working with biologics on large scales is the need for long-term cultures. But even well-established cell lines have limited stability in these conditions. So, you must have proper control systems to ensure the continuity of product supply [4].

For this, you can prepare a pool of cells derived from a single clone and store it in liquid nitrogen in several vials, creating a master cell bank (MCB) [4]. Then you can thaw and expand individual vials as required to create your working cell bank [4].

With this method, you can start thousands of production runs before the original cell stock is exhausted [4].

Needless to say, you must characterize and extensively test this MCB for contaminants such as bacteria, fungi, viruses, and mycoplasmas [4].

Moreover, when doing gene editing experiments, you must ensure that the transfection took place correctly, especially when dealing with large vectors that are more difficult to transfect.

How Can Inish Analyser Help You?

As we discussed earlier, biological products are complex and heterogeneous and must undergo strict quality control.

A reliable cell count and viability are key metrics in biomanufacturing, ensuring:

  • Safety, efficacy, purity, potency, identity, and quality of the final product

  • Oversight of both product and process

  • Quality control of source materials, intermediates, and the final product

  • Reproducibility of lots

  • Comparability after manufacturing change

Another essential metric for cellular genetic engineering applications is transfection efficiency.

And that's precisely what we can help you with. Cellix's Inish Analyser is the most reliable, rapid, label-free instrument for cell counting, cell viability, and transfection efficiency prediction.

With the Inish Analyser, you can predict transfection efficiencies immediately post-transfection by giving you an accurate analysis of the percentage of cells with open membranes.

The main advantages of this method are:

  • Label-freeNo fluorescent dyes, meaning you won't waste time on sample preparation.

  • No expensive slides/chambersSave money for your research.

  • High-throughput - Analyse thousands of cells per second.

  • Compatible with a wide variety of cell samples - CHO cells, T-cells, yeast cells, stem cells, sperm cells, etc.

  • Easy to use and compactRun your samples in three steps.

  1. Pipette 40µL of cell sample into a tube

  2. Add Buffer

  3. Run Sample in Inish Analyser


References

1. Morrow T, Felcone LH. Defining the difference: What Makes Biologics Unique. Biotechnol Healthc [Internet]. 2004 Sep;1(4):24–9. Available from: https://pubmed.ncbi.nlm.nih.gov/23393437

2. Cao J, Perez-Pinera P, Lowenhaupt K, Wu M-R, Purcell O, de la Fuente-Nunez C, et al. Versatile and on-demand biologics co-production in yeast. Nat Commun [Internet]. 2018;9(1):77. Available from: https://doi.org/10.1038/s41467-017-02587-w

3. Walker RSK, Pretorius IS. Applications of Yeast Synthetic Biology Geared towards the Production of Biopharmaceuticals. Genes (Basel) [Internet]. 2018 Jul 6;9(7):340. Available from: https://pubmed.ncbi.nlm.nih.gov/29986380

4. Bird P, Hale G. Cell Banks and Stability of Antibody Production BT - Diagnostic and Therapeutic Antibodies. In: George AJT, Urch CE, editors. Totowa, NJ: Humana Press; 2000. p. 303–7. Available from: https://doi.org/10.1385/1-59259-076-4:303

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