Fluent^® Automation Workstation

Histone acetylation and deacetylation play an important role in the regulation of gene expression. Histone deacetylases (HDACs) are a group of enzymes that remove acetyl groups from ε-N-acetyl lysine amino acids on histones, allowing them to wrap DNA more tightly.This condenses the DNA structure, preventing transcription. Conversely, acetylation neutralizes the positive charges on the histones by changing amines into amides, decreasing their ability to bind DNA.This decreased binding allows chromatin expansion, permitting genetic transcription to take place. 18 HDAC enzymes are currently known, which are either zinc-dependent or nicotinamide adenine dinucleotide-dependent (NAD-dependent).They are grouped into four classes according to their homology to yeast proteins. Classes I, II and IV are considered ‘classical’ HDACs, and are inhibited by trichostatin A (TSA).Due to the involvement of HDACs in various diseases and cancer development, there is now significant interest in selective HDAC inhibitors as potential therapeutic agents.

The failure of preclinical drug entities is a major problem for pharmaceutical companies, accounting for substantial proportions of their precious R&D budgets. As a consequence, there is a need to bring compound toxicity testing into a high throughput environment similar to primary efficacy screening. Metabolism of drugs is a key factor in compound toxicity, and the cytochrome P450 (CYP) superfamily of enzymes – mainly expressed in the hepatocyte cells of the liver – play a very important role in this process. There are now a number of screening assays designed to be automated in a microplate format for medium throughput profiling of compounds which may induce or inhibit CYP activity. In this application note, we describe a 384-well microplate format lead generation method using three different CYP enzymes, illustrating a typical assay run in an ADME laboratory.

For many years, small molecule drug discovery has relied on the evaluation of one compound against another compound.Measurement of the activity of a small molecule against a target must be standardized, and this is achieved by determining the IC50 or EC50 value (half maximal inhibitory/effective concentration respectively) using dose-response curves.To set up dose-response curves, compounds must be dissolved in a solvent to create solutions of varying concentrations, and DMSO has become the established solvent of choice. Today, chemists provide the biologists or compound management teams responsible for producing the dose-response curves with compound solutions prepared in DMSO.The workflow frequently begins with picking the key target compounds from a number of different microplates, generally referred to as hit-picking.These key compounds have often been selected based on single concentration data from a high throughput screening (HTS) campaign.

For many years, small molecule drug discovery has relied on the evaluation of one compound against another compound.Measurement of the activity of a small molecule against a target must be standardized, and this is achieved by determining the IC50 or EC50 value (half maximal inhibitory/effective concentration respectively) using dose-response curves.To set up dose-response curves, compounds must be dissolved in a solvent to create solutions of varying concentrations, and DMSO has become the established solvent of choice.Today, chemists provide the biologists or compound management teams responsible for producing the dose-response curves with compound solutions prepared in DMSO.The workflow frequently begins with picking the key target compounds from a number of different microplates, generally referred to as hit-picking.These key compounds have often been selected based on single concentration data from a high throughput screening (HTS) campaign.

The ability to carry out cell-based G-Protein coupled receptor (GPCR) assays in high throughput is a key part of the drug discovery process.Combining a Fluent laboratory automation solution configured for cell-based assays with the DiscoveRx PathHunter β-Arrestin human and ortholog GPCR assay kits gives users the power to run 384-well plate-based assays in high throughput.This increased processing speed and capacity over current solutions, combined with extremely efficient use of laboratory space, makes this a superior system for cell-based assays.

The NCI-60 panel represents a wide diversity of common cancer-types and it is one of the most important tools in modern drug discovery for the identification of new anti-cancer compounds.Although, the cell lines are commonly used in phenotypic assays for monitoring cell growth and survival, when combined with bioinformatics tools such as the NCI COMPARE database, NCI panel data can also provide a preliminary indication of the potential mechanisms of action for novel compounds.Typically, large sets of compounds are initially tested at a single concentration.Hits showing growth inhibition or cytotoxic properties are profiled in dose response format over multiple cell lines to establish compound potency and selectivity.

Epigenetics has played an important role in drug discovery for many years, but its importance has grown significantly recently due to the role epigenetic modifications can play in cancer and stem cell differentiation. Methylation is the principal epigenetic modification of DNA and of proteins known as histones, which package DNA into chromatin inside a cell.Epigenetic modifications define how genetic information is read and used by cells and, through cell divisions, these epigenetic changes may last for the duration of the cell's life. They may also last for multiple generations, even though they do not involve changes in the underlying DNA sequence of the organism.Epigenetic modifications can therefore be inherited, and are influenced by environmental factors, some of which can induce epigenetic signaling that may contribute to biological processes such as aging.The Cisbio EPIgeneous cell-based assays enable direct measurement of methylation, demethylation, and total proteins on both adherent and suspension cells.Tecan has re-invented automation with Fluent, a unique instrumentation concept built around the applicationspecific needs of your lab. Fluent breaks new ground, delivering more capacity and increased speed.

Epigenetics has played an important role in drug discovery for many years, but its importance has grown significantly recently due to the role epigenetic modifications can play in cancer and stem cell differentiation.Methylation is the principal epigenetic modification of DNA and of proteins known as histones, which package DNA into chromatin inside a cell.Epigenetic modifications define how genetic information is read and used by cells and, through cell divisions, these epigenetic changes may last for the duration of the cell's life. They may also last for multiple generations, even though they do not involve changes in the underlying DNA sequence of the organism.Epigenetic modifications can therefore be inherited, and are influenced by environmental factors, some of which can induce epigenetic signaling that may contribute to biological processes such as aging.The Cisbio EPIgeneous cell-based assays enable direct measurement of methylation, demethylation, and total proteins on both adherent and suspension cells.

Immunosuppressant drugs support solid organ transplantation by preventing rejection by the patient’s immune system. Immunosuppressant levely are tightly monitored to prevent transplant rejection or accumulation of toxic effects.

This application note highlights an example automated sample preparation protocol developed by Tecan for the MassTox® Immunosuppressants in Whole Blood – ONEMinute Test – LC-MS/MS assay kit (Chromsystems) on the Fluent® Mix and Pierce Workstation.

Fluent was designed and built to increase throughput and productivity, enabling laboratories to get more done in less time – and less space.

Designed for those who ‘press go’, daily activities such as running assays and routine maintenance become as straightforward as using the smartphone in your pocket. Users intuitively know what to do next.

Fluent Gx brings the proven performance enhancements of the Fluent Laboratory Automation Solution to highly regulated laboratories.Application workflows in clinical and other regulated areas can benefit from high throughput, ease of operation and outstanding precision & accuracy with maximum process security.

Tecan’s groundbreaking Fluent platform brings unsurpassed speed and flexibility to plate formatting and copying. Throughput and accuracy are everything, so we’ve engineered a compact automation solution with three arms that work in parallel across a Dynamic Deck™ offering unmatched plate capacity. Discover an instrument that speaks the language of your laboratory, making compound management more simple, efficient… and fluent.

Seamless integration of cell-based assay technologies streamlines your workfl ows and simplifi es your laboratory life.

Fluent’s groundbreaking design delivers more capacity and increased speed for genomic workflows. This innovative solution offers superior precision, higher throughput and longer walkaway times. Get more done in less time, with greater confidence.

The MultiChannel Arm™ 384 has been designed to offer higher productivity to automated liquid handling workflows in pharmaceutical and biotechnology research applications, increasing the efficiency and speed of pipetting processes, as well as delivering an outstanding level of flexibility.

For Freedom EVO® and Fluent®

Accurate nucleic acid quantification and quality control without sample loss

Module for Fluent® automation workstation specifications and typical performance values

Whole blood screening is performed regularly for patients taking drugs that are absorbed into blood cells, such as immunosuppressants prescribed after organ transplants. However, sample preparation is challenging for laboratories using LC-MS for drug quantification.

The Fluent® Mix and Pierce Workstation enables automated processing of immunosuppressant kits, providing consistency and accuracy. This brochure highlights an example automated sample preparation protocol developed by Tecan for the MassTox® Immunosuppressants in Whole Blood – ONEMinute Test – LC-MS/MS assay kit (Chromsystems).

The Fluent Gx Mix and Pierce Workstation enables users to develop workflows in forensic investigations. As crime scene samples are scarce and cannot be replaced, any loss of material needs to be avoided.

The Fluent Gx Mix and Pierce Workstation minimizes the risk of sample loss by adding extraction solvents directly to the sample collection tubes, and mixing in a controlled manner. This flyer highlights an example workflow for non-biological sample material.

The Multiple Channel Arm™ 96 (MCA 96) for the Fluent® liquid handler offers an incredible volume range ideal for high-throughput workflows.

Fluent specifications.

Enhanced capacitive liquid level detection for increased reliability and reproducibilitySpeed and throughput seem like logical reasons to acquire an automated liquid handling system but, for many users, the consistency of results is an even more valuable asset.An unobtrusive, barely noticed feature – liquid level detection – plays a critical role in ensuring the reliability and consistency of the pipetting processes, but not all liquid level sensing technologies are created equal.Tecan’s Adaptive Signal Technology is a new generation of capacitive liquid level sensing that does far more than just detect the liquid level in a microplate well; it can reliably distinguish between the true liquid level and bubbles, detect tip occlusions, prevent wells over-flowing, and offer new opportunities for in-process error handling.

Reducing the liquid dead volume is a highly desirable aspect of a liquid handling platform’s performance, particularly in the case of expensive reagents or precious scarce samples.Minimizing the dead volume not only presents substantial reagent and sample cost savings, it can also result in more assays per unit volume, further decreasing labor costs and allowing more data per cycle.Fluent Laboratory Automation Solutions with Adaptive Signal Technology™ (AST) advanced liquid level detection offer several synergistic benefits that reduce dead volume without compromising on the precision or accuracy of liquid transfers.