Effective Ways to Reduce Cell-Based Assay Development Time

The successful development and use of cell-based assays can have a huge impact on the cost-effectiveness of pharmaceutical research. Evaluating the biological effects of chemical compounds requires multiple different types of approaches, including molecular, cellular, and biochemical. The first step towards such evaluation includes the successful execution of cell-based and in vitro assays.

The assay readouts thus obtained are relevant to disease identification, human health, therapeutic methodologies, and drug development. Reduction in the time required for the development of cell-based assays can make the process of drug discovery much cheaper and more efficient. Late-stage failures in the process of drug development can cause heavy financial losses as well as massive delays in the production process.

Understanding disease biology and defining the associated target for disease control are the first steps in the process of drug discovery. During the development phase of the assay, multiple validation steps are performed. This can be time-consuming because failure to satisfactorily validate the assay can lead to distorted results, which would hamper the drug development process as a whole.

The discovery and development of new chemical compounds (such as medicines and drugs) can be very expensive, with the costs rising sharply through the successive stages of development. Some of how the development time of a cell-based assay can be reduced, to minimize the mounting costs and prevent late-stage failures, have been mentioned below.


  • Temperature Control

Stable temperatures are essential for the execution of a reliable cell-based assay, especially when assay kinetics in living creatures is being studied. Most assay protocols specify the need for ‘ambient temperature’, which means that any thermal variations in the lab environment or within the assay instrument can compromise the reliability and quality of the assay.

Motors and other moving fragments within a microplate reader can generate a significant amount of heat, which could potentially harm the reproducibility of a cell-based assay. A microplate with numerous readers might also cause variability and poor precision, due to the occurrence of temperature gradients. A plate reader that enables active temperature management can help overcome some of these challenges.


  • Physiological Optimization

Cell-based assay development requires proper physiological optimization within relevant parameters. The correct physiological mechanism is essential for determining the optimal reagent concentration in a cell-based assay. Using only statistical parameters in assay optimization can, therefore, be counterproductive, because it does not take into account the physiological aspects of the process.

The salt (or reagent) concentration required for perfect reproducibility in an in-vitro assay might, for instance, be completely different from the environment that a cell would contend with inside the human body. This is why it is important to keep physiological factors and conditions in mind when using statistical parameters and performance metrics for assay optimization.


  • Fluorescence-Based Assays

In cell-based assay development, fluorescence techniques are much more prevalent than absorbance-based methods, particularly when the microtiter plate format is being used to conduct the assay. The two primary reasons for these are the short path-length typical in miniaturized assays and the impact of this path-length on signal strength. As a result of these factors, miniaturized absorbance-based assays often lose their signal window.

One major issue with fluorescent assays is that they might face interference from light-absorbing compounds in the emission or excitation range of the assay. On the other hand, if the compounds themselves are fluorescent, this could give rise to the problem of a false negative. Overcoming these challenges can go a long way in reducing the amount of time needed for cell-based assay development.


  • Interference Minimization

Red-shifted fluorophores, which possess longer wavelengths, are often used in cell-based assays to minimize compound interference; but these require a subtle detection instrument comprising a broad spectral range to work. A multimode reader with enhanced PMTs optimized for far-red dyes can be used for superior cell-based assay performance.

There are multiple – and often unpredictable – ways in which compounds of interest can affect a cell-based assay. Autofluorescence and quenching are the two primary mechanisms of direct interference of a compound with an assay. Hence, it is important during an assay to have an orthogonal method for confirming activities not subject to interference by either of these mechanisms.


  • Flexibility and Sensitivity

In the process of assay miniaturization, the gap between sensitivity and flexibility needs to be bridged for the process to be a success. Since miniaturization also helps reduce the consumption of expensive assay reagents, this also increases the cost-effectiveness of the drug-development process. Lowered sensitivity and changes in the surface-to-volume ratio are some of the challenges that researchers often encounter during the miniaturization process.

Every reduction in volume leads to a simultaneous reduction in the sensitivity of the assay because fewer product-sensing materials can be added. Therefore, a suitable detector needs to be found for the changes in pH levels and the resultant shifts in detectable wavelength peaks. Such a detector would help create a balance between the required amount of flexibility and sensitivity in a cell-based assay.


  • Workflow Automation

Assay automation can reduce the risk of human error while speeding up routine operations, resulting in faster and more efficient statistical data output. The use of microplate stackers for improved reliability in routine applications, the reduced time needed for analysis, and automated batch processing for greater productivity in the laboratory is also quite common.

Typical workflow steps like restacking and plate-loading can be automated through these microplate stackers. Intelligent software design can facilitate the automatic scheduling of operations based on certain readouts and assay conditions. Such software can make it easy to set up almost any measurement protocol that would be beneficial in fostering cell-based assay development.


In Conclusion

While the above-mentioned steps can help reduce the amount of time required for cell-based assay development, thus making the process of drug development more cost-effective than ever before, it is essential to only entrust a reputed lab with the task of performing cell-based assays. Such labs have the required technology and manpower to perform accurate, reliable, and timely cell-based assays for high-quality research.