Protein engineering is an important discipline within synthetic biology and involves the generation of optimized proteins for many applications. This can include improving the amino acid sequence or modifying the protein to improve function. Researchers can also integrate novel synthetic amino acids into their proteins to optimize specific characteristics1.
Proteins are core functional components of biological systems, making protein engineering applications virtually limitless in healthcare, agriculture, and environmental protection applications (Fig. 1)2. Deciding what changes to make to a protein is a problematic initial step; however, the practical aspects of protein engineering workflows also pose several challenges.
Figure 1. Proteins are complex molecules that must fold properly to achieve the desired functionality. (Source)
Precise liquid handling is essential for accurate and efficient protein engineering. The I.DOT Liquid Handler combines non-contact dispensing with low-volume capabilities to provide speed and scalability to protein engineering workflows. In this blog, we explore the importance of accurate liquid handling in overcoming challenges in protein engineering workflows.
Proteins are capable of folding and interacting with their environment in ways that are difficult to predict, even with advanced prediction software3. This means that reaction mixtures for protein engineering must be tightly controlled and coupled to a technology that can handle precise volumes and dispense them accurately. Minute inaccuracies can lead to improper folding and assembly, significantly impacting downstream functionality.
Reactions in protein engineering are sensitive to pH, temperature, and reagent concentrations4. Slight changes in these parameters due to inaccurate liquid dispensing can significantly disrupt protein engineering reactions and lead to reduced yield, incorrect sequence or modification, or complete workflow failure.
Furthermore, reagents for protein engineering can be expensive, which means that failed workflows carry a significant risk in terms of resources. Ultimately, these challenges can lead to slower research output, which can make the difference between securing a grant or being the first to gain market approval for a novel therapy.
Protein engineering workflows must be highly controlled and optimized to achieve the best results. Researchers must consider many different parameters of reagent dispensing to maximize accuracy and throughput. These parameters include:
Proper equipment calibration ensures that the desired liquid volume is being dispensed5. This is essential when following vendor protocols and eliminates an important variable when troubleshooting. Maintaining proper calibration is also important for ensuring reproducibility of results across long periods of time. If a protocol suddenly stops working or protein yield decreases over time, lack of calibration is a likely culprit.
2. Tip Selection
Selecting the correct pipette tip (or completely omitting pipette tips) ensures accuracy and efficiency in protein engineering workflows. Low retention tips are helpful when pipetting protein samples, and some suggestions are designed to reduce shear stress, which proteins are susceptible to. Choosing the appropriate pipette size for the volumes you are using is also essential.
Dispensing speed and height can negatively impact reaction efficiency6. Proteins in solution are sensitive to unfolding if they are not dispensed correctly. Most researchers will remember a time when their entire solution turned to foam because they pipetted it too quickly. Choosing the optimal dispensing height is vital for avoiding splashing and using surface tension effects (Fig. 2) to achieve slow dispersal of reagents.
Figure 2. Surface tension effects are important in accurate dispensing, especially in microfluidics systems. (Source)
Reducing dead volume (i.e., the volume left in the tip after dispensing) is important for resource efficiency. Even tiny volumes lost during each dispensing can add up to a significant resource loss over time. The I.DOT Non-Contact Dispenser achieves a dead volume of 1 µL, which is far lower than other dispensers.
Choosing multichannel dispensing allows researchers to perform multiple reactions in parallel. The I.DOT Liquid Dispenser possesses this ability, significantly increasing throughput and reducing turnaround times in protein engineering workflows.
Optimized liquid handling offers many advantages to researchers performing protein engineering workflows. Technologies like the I.DOT Non-Contact Dispenser help to ensure accurate and reproducible protein engineering experiments. The machine can be programmed to repeatedly execute a precise protocol, giving researchers confidence in their experiment even over long periods of time. Technologies like this also offer scalability and reduced dead volume, meaning researchers save money and can use fewer reagents than those stated by vendors without sacrificing quality.
Finally, automated liquid handling significantly increases the efficiency of protein engineering experiments by achieving both accuracy and high throughput7. In today's competitive environment, combining faster turnaround times with high-quality output is essential to gaining an advantage over competitors and establishing leadership in a given industry.
Optimizing liquid handling is crucial for enhancing accuracy, efficiency, and reproducibility in protein engineering workflows. These improvements save resources and accelerate research, giving scientists a competitive edge in developing innovative solutions in healthcare, agriculture, and environmental protection.
Mastering protein engineering requires meticulous control. The I.DOT Liquid Handler by DISPENDIX delivers unmatched accuracy with non-contact dispensing as low as 4 nL, minimizing errors and maximizing your yield. Its integrated droplet detection ensures every transfer is successful, preventing wasted precious proteins and reagents.
Optimize your protein engineering workflows and see groundbreaking results. Book a demo today to learn more about the I.DOT Liquid Handler!