Screening
Figure 20
IonWorks systems. With five IonWorks platforms worldwide, it has the capacity to conduct large electrophysiology screens (up to 50K samples) with high quality and fast turnaround for both voltage and ligand-gated targets. In parallel, Essen can sup- port multiple SAR and safety profiling campaigns as well as high throughput mechanistic analyses. Our higher throughput approach couples the pre- cision and relevance of translational electrophysi- ology protocols with an affordability that permits early and wider scale testing (eg for cardiac safety assays such as hERG and hNaV1.5). Essen has also adopted a multitude of business relationships to meet the needs of its clients ranging from fee-for- service to collaborations involving structured mile- stone payments (Figure 19).
Fluxion Bioscience’s (
www.fluxion.com) IonFlux system applies well plate microfluidics technology to ion channel screening. This next generation auto- mated patch clamp instrument is the only screening system to include continuous compound perfusion and continuous recording during the entire protocol. This combination enables superior assay flexibility and key advantages for ligand-gated screens, where throughput is increased by an order of magnitude over other automated systems. The use of intercon- nected microfluidic channels that are coupled to a pneumatic interface removes the need for liquid han- dling; the unit operates much like a plate reader and can be used either stand-alone or coupled to stan- dard liquid handlers where more throughput is required. Another unique feature is the available heating module, which will be improved by the upcoming introduction of a chiller plate which can also lower recording temperatures to 4°C. The mod- ular nature of the instrument allows for multiple readers attached to the same liquid handling robot to scale up assay throughput easily. Recent customer data from NMDA and Ach receptors highlights the advantages of simultaneous compound application and efficient washout, achieving very high (>95%)
Drug Discovery World Fall 2011
success rates for these difficult ligand-gated ion chan- nel targets. While the Fluxion’s current microfluidic consumable plate records currents from 20-cell ensembles, a single-cell recording plate, called the F1, will be introduced in Q4 2011. The F1 plate includes gigaohm seal resistance capability, and will bolster the effectiveness of the IonFlux for characterising heterogenous cell populations and other applications requiring single-cell recordings or gigaohm seals. The F1 single-cell recording plate is fully compatible with existing IonFlux systems (Figure 20).
In June 2011 flyion (
www.flyion.com) released its f11™ series of automated patch clamp instruments which will succeed all previously offered products. The f11 has been co-developed with a medical devices manufacturer in Switzerland. It is assembled in an ISO 13485 certified production environment as a GLP compliant device. The f11 is based on the patented Flip-the-Tip™ technology which performs the patch clamp experiment inside a regular glass pipette. Instead of pressing a micropipette against a cell membrane and applying suction to form a high resistance seal between the glass and the cell mem- brane, a cell suspension is infused into the micropipette and suction is applied from the aper- ture until the cell seals the pipette from the inside.
Figure 21 flyion’s f11™ – the only commercially available automated patch clamp instrument using glass pipettes
Schematic of the microfluidics for one experimental pattern of Fluxion’s IonFlux system. A main channel intersects trapping regions containing intracellular solution that capture cells at the ensemble recording array. All compound channels converge just upstream of the 20-cell recording array. Inset left: Micrograph of an ensemble recording array composed of 20 microfluidic channels occupied by captured cells. Right: A cumulative dose response consisting of four compound applications (three agonist concentration and wash buffer) injected across the entire consumable (64 recording channels, one quadrant shown) to obtain 384 responses to compound additions in a space of 20s
55
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60 |
Page 61 |
Page 62 |
Page 63 |
Page 64 |
Page 65 |
Page 66 |
Page 67 |
Page 68 |
Page 69 |
Page 70 |
Page 71 |
Page 72 |
Page 73 |
Page 74 |
Page 75 |
Page 76 |
Page 77 |
Page 78 |
Page 79 |
Page 80 |
Page 81 |
Page 82 |
Page 83 |
Page 84 |
Page 85 |
Page 86 |
Page 87 |
Page 88 |
Page 89 |
Page 90 |
Page 91 |
Page 92