May 15, 2011 (Vol. 31, No. 10)

The SBS meeting marked the U.S. launch of Reinnervate’s alvetex® cell culture scaffold for 3-D in vitro cell growth. Initially marketed in Europe in a 12-well plate format, alvetex is now available as 6-well and 12-well inserts. It is supplied as a gamma-irradiated, ready-to-use polystyrene membrane and can be optimized for specific cell types and applications. Users can control the degree of penetration within the 3-D structure and optimize the culture format and parameters to support long-term cell culture. The new well inserts allow cells to be fed simultaneously from above and below. They fit into multiwell plates or into insert holders designed to fit into a Petri dish.

Jens Kelm, head of product management and co-founder of InSphero, presented a tutorial on the benefits of using 3-D microtissue models for screening to evaluate drug efficacy and toxicity. InSphero developed an assay-ready microtissue mass production technology that generates 3-D microtissues of various tissue types containing 100–5,000 cells, without the use of a scaffold. Customers can purchase standard microtissues or provide InSphero with their own cell lines. The company recently introduced tumor microtissue models for colon, liver, prostate, and kidney cancer.

The Automation Partnership recently announced a name change to TAP Biosystems. The company also introduced a 96-well version of its RAFT™ (Real Architecture for 3D Tissue™) system for generating multi-cellular 3-D collagen-based tissue models.

Kuraray’s  micro-space cell culture plate designed to support 3-D cell growth is currently available in Japan and will be introduced to the European and U.S. markets in early to mid-2012.

HepG2 cells cultured for two weeks on Reinnervate’s alvetex® inserts (magnification x100).

Cellular Image Analysis

GE Healthcare introduced the IN Cell Analyzer 6000 high-content cellular-analysis system, which features a variable iris-like aperture, laser-based confocal system that is fully adjustable and controlled via a user interface to maximize speed, image quality, and performance channel-by-channel for a particular assay. It can range from widefield to full confocal imaging. The new Investigator 2.0 software platform supports the IN Cell Analyzer 6000.

Genedata Screening® 9, new from Genedata, features an enhanced user interface that allows for simultaneous processing of multiple dose-response assays and new display capabilities for high-content screening that enable users to view images and accompanying data together, making it easier to identify outliers and to streamline the process of going from plate to well to image. Images are generated as an experiment is under way, allowing users to visualize the kinetics and optimize assay parameters.

Epigenetic Strategies

Promega featured oncology-related drug discovery tools including its ADP-Glo™ Max homogeneous luminescent assay for measuring ATPase or kinase activity. The company’s GSH/GSSF-Glo™ assay quantifies total glutathione, GSSG levels, and GSH/GSSG ratios in cultured cells as a measure of oxidative stress and an indicator of a toxicologic drug response. The bioluminescent histonedeacetylase (HDAC) assay is designed for cell-based screening of HDAC enzyme activity and potential inhibitors of these epigenetic modifiers.

PerkinElmer added 12 new reagents to its AlphaLISA® platform for epigenetics research. The homogeneous proximity immunoassays measure methylation or acetylation and can accommodate full-length histones and nucleosomes.

Screening Technology

Horizon Discovery’s virally mediated Genesis™ genome-editing technology enables targeted engineering of endogenous genes in human cells lines to create genetically defined knock-out or knock-in cellular models. Homology cassettes carried in recombinant adeno-associated virus (AAV) vectors are used to perform targeted homologous recombination, ensuring no off-target integrations or sequence errors.

Horizon has applied this technology to create X-Man™ patient-relevant human cell lines for personalized medicine applications, designed to model the disease-causing mutations present in a patient’s tumor. These patient-specific models can be used to study drug activity, therapeutic response, and drug resistance, and to design individualized treatment regimens and select patients for clinical studies.

The company recently joined the European Union Colon Cancer and Therapeutics (COLTHERES) consortium and is generating X-Man colon cancer cell lines that incorporate predictive biomarkers of drug response and resistance, as well as diagnostic and prognostic biomarkers.

Johannes Ottl, unit head of label-free technologies at Novartis Pharma, presented a poster entitled, “SPR in early phases of low molecular weight lead discovery,” in which he described the use of surface plasma resonance (SPR) in screening campaigns for hit identification and for validation and characterization of binding, including affinity and kinetic measurements, of low molecular weight compounds to a target.

Ottl’s group is using chip-based SPR technology from Bio-Rad that allows for screening multiple protein samples in parallel, which he notes, offers advantages for running replicates and evaluating compound selectivity and modified/mutated proteins side-by-side in the same experiment.

Bio-Rad’s ProteOn™ XPR36 Protein Interaction Array System allows users to immobilize a panel of 6 ligands on the surface of a 6×6 chip and to measure 36 interactions at one time. The chip surface is composed of a modified alginate polymer layer bound to the gold surface of the sensor prism. Bio-Rad recently introduced the ProteOn HTG sensor chip, containing tris-NTA complexes immobilized to the gold surface to capture His-tagged proteins.

The SoPRano™ SPR-based label-free high-throughput screening platform from Pharma Diagnostics designed for performing protein binding assays, enables localized SPR detection on gold nanoparticles and is compatible with absorbance at 530 nm on standard plate readers. The initial SoPRano kit utilized passive charge absorption coupling chemistry. The company recently launched kits with biotin-direct coupling chemistry and carboxyl covalent coupling for stable conjugation of amine-containing molecules.

HighRes Biosolutions’ new TCell™ table-based automation system offers a variety of enclosure options, including BSL2, HEPA filtration, laminar flow, and temperature and humidity control. It contains three-axis device platforms for ease of access to instruments, the Cromium robot powered by a harmonic drive gearing system, under-table storage, and an optional MicroDock for quick switching of additional devices such as readers, incubators, or pipettors.

The Picodrop 200 spectrophotometer from Micronic North America  performs protein or DNA quantification in 2 µL samples in a patented disposable pipette tip with a three-second read time. The device uses a UV xenon light source and can read across a wavelength range of 220–950 nm.

Later this year, a collaborative venture between Tecan and HP will bring to the drug screening market dispensing products based on HP’s inkjet technology.

According to Horizon Discovery, X-Man (mutant and normal) cell lines are available that model many of the common genotypes observed in human cancer, plus provide their perfectly matched normal genetic backgrounds as a reference.

Assay Technology

BioFocus and Almac initiated a collaboration intended to provide BioFocus customers access to compound screening and profiling services using Almac’s Flexyte™ fluorescence lifetime, antibody-free, homogeneous assays. In addition to Flexyte assays for protein kinase and protease targets, Almac has introduced assays for epigenetic targets including protein methyltransferases and deiminases, designed to probe the interactions between epigenetic enzymes and histone proteins.

For high-throughput genomic applications, Luminex developed qBead™-based gene-expression assays that combine htg Molecular’s  quantitative nuclease protection assay (qNPA™) chemistry with Luminex’ xMAP® MagPlex® bead-based multiplex technology.

TaqMan® OpenArray® MicroRNA Panels from Life Technologies combine TaqMan miRNA assays with the nanofluidic OpenArray Real-Time PCR platform to achieve high-throughput global screening of human miRNAs. The panels provide for the generation of more than 27,000 miRNA data points in an eight-hour period, screening 754 unique human miRNAs in up to 36 samples, according to the company.

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