Automated liquid-handling operations for robust, resilient, and efficient bio-based laboratory practices

Torres‐Acosta, Mario A.; Lye, Gary J.; Dikicioglu, Duygu

doi:10.1016/j.bej.2022.108713

Cited by 26 publications

(14 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To perform qPCR, cells must be lysed for RNA extraction and therefore do not require the use of an optimized cell digestion protocol which preserves whole cells. [ 53 ]…”

Section: Resultsmentioning

confidence: 99%

An Automation Workflow for High‐Throughput Manufacturing and Analysis of Scaffold‐Supported 3D Tissue Arrays

Cao

Latour

et al. 2023

Adv Healthcare Materials

View full text Add to dashboard Cite

Patient‐derived organoids have emerged as a useful tool to model tumour heterogeneity. Scaling these complex culture models while enabling stratified analysis of different cellular sub‐populations, however, remains a challenge. One strategy to enable higher throughput organoid cultures is the scaffold‐supported platform for organoid‐based tissues (SPOT). SPOT allows the generation of flat, thin, and dimensionally‐defined microtissues in both 96‐ and 384‐well plate footprints that are compatible with longitudinal image‐based readouts. SPOT is currently manufactured manually, however, limiting scalability. In this study, an automation approach to engineer tumour‐mimetic 3D microtissues in SPOT using a liquid handler is optimized and comparable within‐ and between‐sample variation to standard manual manufacturing is shown. Further, a liquid handler‐supported cell extraction protocol to support single‐cell‐based end‐point analysis using high‐throughput flow cytometry and multiplexed cytometry by time of flight is developed. As a proof‐of‐value demonstration, 3D complex tissues containing different proportions of tumour and stromal cells are generated to probe the reciprocal impact of co‐culture. It is also demonstrated that primary patient‐derived organoids can be incorporated into the pipeline to capture patient‐level tumour heterogeneity. It is envisioned that this automated 96/384‐SPOT workflow will provide opportunities for future applications in high‐throughput screening for novel personalized therapeutic targets.

show abstract

“…To perform qPCR, cells must be lysed for RNA extraction and therefore do not require the use of an optimized cell digestion protocol which preserves whole cells. [ 53 ]…”

Section: Resultsmentioning

confidence: 99%

An Automation Workflow for High‐Throughput Manufacturing and Analysis of Scaffold‐Supported 3D Tissue Arrays

Cao

Latour

et al. 2023

Adv Healthcare Materials

View full text Add to dashboard Cite

show abstract

“…The SPRI bead purification procedure requires only basic laboratory instruments, reaction tubes, pipettes, and magnets, [15,16] thus, this procedure is directly compatible with automation where minimal manual interference is required. [23] In contrast, methods that are based on precipitation or filtration rely on centrifugation [3,14,21,22] which is a significant obstacle for automation. Besides the automated liquid handling robot we used here (Biomek NX P Automated Workstation), various models of automated liquid handling robots can be found on the market that provide either full automation or partial automation including the chemagic Prepito from Revvity, KingFisher from Thermo Fisher Scientific, and Assist Plus from Integra, alternatively, liquid handling machines can be made with custom 3D printed parts.…”

Section: Resultsmentioning

confidence: 99%

“…[15][16][17][18][19][20] In contrast to other methods, this technique does not require centrifugation [3,14,21,22] or modifications of the origamis, which present limitations for automated large-scale implementations, and we demonstrate the readiness of this method for scaling up by performing an automated purification of 96 DNA origami samples with a liquid handling robot. [23] We envisage that the SPRI clean-up method will further the commercial exploitation of DNA nanostructures by enabling their high-throughput purification. The DNA origamis are mixed with the SPRI beads at a specific beads-to-sample volume ratio, followed by their separation with an external magnet.…”

Section: Introductionmentioning

confidence: 99%

Automated Purification of DNA Origami with SPRI Beads

Chau,

Mohanan,

Macaulay

et al. 2023

Small

View full text Add to dashboard Cite

DNA origami synthesis is a well‐established technique with wide‐ranging applications. In most cases, the synthesized origami must be purified to remove excess materials such as DNA oligos and other functional molecules. While several purification techniques are routinely used, all have limitations, and cannot be integrated with robotic systems. Here the use of solid‐phase reversible immobilization (SPRI) beads as a scalable, high‐throughput, and automatable method to purify DNA origami is demonstrated. Not only can this method remove unreacted oligos and biomolecules with yields comparable to existing methods while maintaining the high structural integrity of the origami, but it can also be integrated into an automated workflow to purify simultaneously large numbers and quantities of samples. It is envisioned that the SPRI beads purification method will improve the scalability of DNA nanostructures synthesis both for research and commercial applications.

show abstract

“…With the prevalence of multi-omics and large cohort studies, automating sample handling is essential in academic and clinical research. Because of the increased throughput and reproducibility, while simultaneously reducing contamination, automated liquid handlers hold a highly attractive advantage [19,20]. Thus, we developed an automated protocol for enzyme immobilization and subsequent digestion with multiple silicone-based linkers, and enzymes to leverage these bene ts.…”

Section: Introductionmentioning

confidence: 99%

Development of Automated Proteomic Workflows Utilizing Silicon-Based Coupling Agents

Frey,

Arad,

et al. 2024

Preprint

View full text Add to dashboard Cite

Automated methods for enzyme immobilization via 4-triethoxysilylbutyraldehyde (TESB) derived silicone-based coupling agents were developed. TESB and its oxidized derivative, 4-triethoxysilylbutanoic acid (TESBA), were determined to be the most effective. The resulting immobilized enzyme particles (IEPs) displayed robustness, rapid digestion, and immobilization efficiency of 51 ± 8%. Furthermore, we automated the IEP procedure, allowing for multiple enzymes, and/or coupling agents to be fabricated at once, in a fraction of the time via an Agilent Bravo. The automated trypsin TESB and TESBA IEPs were shown to rival a classical in-gel digestion method. Moreover, pepsin IEPs favored cleavage at leucine (>50%) over aromatic and methionine residues. The IEP method was then adapted for an in-situ immobilized enzyme microreactor (IMER) fabrication. We determined that TESBA could functionalize the silica capillary's inner wall while simultaneously acting as an enzyme coupler. The IMER digestion of bovine serum albumin (BSA), mirroring IEP digestion conditions, yielded a 33-40% primary sequence coverage per LC-MS/MS analysis in as little as 15 minutes. Overall, our findings underscore the potential of both IEP and IMER methods, paving the way for automated analysis and a reduction in enzyme waste through reuse, thereby contributing to a more cost-effective and timely study of the proteome.

show abstract

Automated liquid-handling operations for robust, resilient, and efficient bio-based laboratory practices

Cited by 26 publications

References 47 publications

An Automation Workflow for High‐Throughput Manufacturing and Analysis of Scaffold‐Supported 3D Tissue Arrays

An Automation Workflow for High‐Throughput Manufacturing and Analysis of Scaffold‐Supported 3D Tissue Arrays

Automated Purification of DNA Origami with SPRI Beads

Development of Automated Proteomic Workflows Utilizing Silicon-Based Coupling Agents

Contact Info

Product

Resources

About