SURFE²R 96SE - ハイスループット・トランスポーター評価系

SURFE²R N1 同様、SURFE²R 96SE は微小なトランポーター/ポンプ電流を検出するためにSSM法による電気生理学を採用し、スループットは96ウェル同時測定へと向上させています。トランスポーターサンプルは 96ウェルフォーマットの金コートのセンサーに適用します。ユーザーの負担をさらに低減するため、すべてのコーティング行程を SURFE²R 96SE 本体で実施することが可能です。どのような基質もトランポーターの活性化に使用できます。The resulting transport or binding currents will be detected and analyzed accordingly. サンプル調製、測定、データ解析に至るまでの全プロセスの完全自動化が可能です。

SURFE²R 96SE の特徴

• 低ノイズの96chアンプによる96ウェル同時測定
• 10,000 データポイント / 日
• 小型自動分注機 CyBio^® Felix に統合
• サンプル調製を含む実験ワークフローを自動化
• EC₅₀等のデータ解析、エクスポートを全自動化
• 8 positions for different solutions (1 well to 96 well)

SSM法による電気生理学の特徴

• 100以上のターゲットバリデート済み、SSM-電気整理学を使用した100以上の査読付き論文
• トランスポーター、ポンプ、リガンド依存性チャネル
• 非起電性トランスポーターや糖類の活性評価にも適用可能
• 精製したトランポーター、細胞から調製したベシクル、プロテオリポソームなど
• ラベルフリーの電気的測定
• 0.1～1 µg タンパク/センサーの少サンプル量で100実験まで実施可能
• 5 msの高分解能のリアルタイムデータ取得（シングルポイントの検出ではありません）
• パッチクランプ法より高いシグナル増幅率
• 律速な結合キネティクスの評価
• EC₅₀、IC₅₀、速度定数、トランスポーター遺伝子多型の比較など

当測定技術では、1990年後半に確立した Solid-Supported Membrane- (SSM法) にも土尽く電気整理学測定手法を採用しています。SSM-電気整理学測定法の詳細は Technology section に示しています。SURFE²R 96SE は SURFE²R N1 以上のハイスループット評価が必要な化合物またはトランポーターのスクリーニング・プロジェクト、スループットを必要とする基礎研究においてに最適なプラットフォームです。SURFE²R 96SE はSURFE²R N1より約100倍の生産性を誇っています。

詳細情報：

SURFE²R 96SE 装置 & ソフトウェア

The SURFE²R 96SE device

The SURFE²R (surface electrogenic event reader) technology is the only available commercial solution for SSM-based electrophysiology on the market. Nanion Technologies developed the SURFE²R N1 as an easy-to-learn all-in-one device for universities and basic research and the high-throughput device SURFE²R 96SE.

The SURFE²R 96SE is integrated into the CyBio^® Felix liquid handling platform. Nanion Technologies provides the electrophysiological hardware and the computer running a software package for data recording and analysis. The device can not only be used for electrophysiological measurements, but also for sample preparation and pipetting purposes. The device can measure 10,000 data points a day in a fully automated manner. In contrast to the SURFE²R N1, the SURFE²R 96SE also comes with a dedicated data analysis software which allows for automated analysis of the massive amount of data you generate each day.

The SURFE²R 96SE uses sensors in a standard 96-well plate format. Membranes containing the protein of interest are adsorbed onto these sensors. The measurement is initiated by a substrate concentration jump. The solution exchange is controlled by the liquid handling platform CyBio^® Felix. The pipetting head contains 96 pipettes which load solutions from solution positions and inject them to the sensor plate. All 96 sensors of the sensor plate are measured simultaneously, but different solutions or transporters can be used for each measurement.

The sensor plate is enclosed by a Faraday cage and connected to a 96-channel amplifier as well as 96 gold pins which work as reference electrodes. The CyBio^® Felix holds up to 8 well plates for 8 to 8 x 96 different solutions which allows up to 7 sequential measurements using different conditions in each automated workflow. The CyBio^® pipettor allows the measurement of fast kinetics with low solution consumption, showing a time resolution of down to 5 ms by using only 50 µl of compound solution per measurement.

Software Package

Our software department developed a software package exclusively for the SURFE²R 96SE.

• The SurfControl works as the recording software and displays the data in real-time
• DataControl works as the analysis software and can be configured to automate your data analysis
• In addition, the CyBio Composer plus pre-installed workflows operate the liquid handling platform and
communicates with the recording software

消耗品

SURFE²R 96SE センサープレート

SURFE2R 96SE Sensor Plate

SURFE²R 96SE センサープレートは，SURFE²R 96SE専用に開発された革新的なNanion Technologiesの自社製品です。製品はパートナー会社で製造後，ドイツのナニオン本社ラボで厳格な品質チェックを行い，ドイツミュンヘンから世界のユーザーへ発送されます。

Material

The core structure of SURFE²R 96SE sensor plates are gold-coated sensors with 3 mm diameter. This structure is incorporated into a 96 well plate for easy handling. The sensor plate is enclosed by a Faraday cage and connected to a 96-channel amplifier during the experiment.

SURFE²R 96SE センサープレート製品ラインナップ

"SURFE²R 96SE Sensor Plate": 96-well recording plate for the SURFE²R 96SE (Order # 182001)

評価済みターゲット: SSM- 電気生理学的測定

インタビュー & ケーススタディ

Analytik Jena - Maria Barthmes about the Cooperation on the SURFE²R Technology

Icon N1 ““We have tested several liquid-handling platforms. CyBio FeliX was the best choice for our application.”

Dr. Maria Barthmes, Product Manager SURFE²R 96SE
Nanion, Munich

Download the full case study here:

Icon N1 SURFE²R Technology Case Study (2020) (1.4 MB)
Transporter Protein Research Redefined

preview the file here for download

Back

Dr. Thomas Seeger & Karin V. Niessen - Statement about the SURFE²R Technology

Icon N1 “The SURFE²R makes extremely challenging electrophysiological targets accessible to robust routine analyses. For example, we have developed an assay for investigating nicotinic acetylcholine receptor ion channels, using membranes from the Pacific electric ray, Torpedo California. In this way, we overcome many of the well-known difficulties associated with this ion channel, which allows us to efficiently obtain information regarding compound pharmacology. Obtained pharmacology values match those of patch clamp recordings exceptionally well. We find that the SURFE²R is an excellent platform for characterization of electrogenic processes in isolated membranes.”

Karin V. Niessen, Dr. Thomas Seeger
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany

Prof. Dr. Klaus Fendler and Co-Authors - Interview about the SURFE²R Technology

Icon N1 ““SSM-based electrophysiology is a suitable technique to study bacterial ion-translocating membrane- systems which are otherwise difficult to approach using conventional electrophysiological techniques, like patch clamp.
Additionally, the mechanical stability of SSMs facilitates the transport activation through fast solution exchange, a condition that is technically challenging in a setup of a bilayer separating two solution-containing compartments.“

Dr. Miyer Fabián Patiño-Ruiz,
Faculty of Science and Engineering, University of Groningen, the Netherlands

Dr. Andre Bazzone, Application Scientist for the SURFE²R product family interviewed the authors of the publication:
"Mutation of two key aspartate residues alters stoichiometry of the NhaB Na⁺/H⁺ exchanger from Klebsiella pneumoniae"
Published 2019 in Nature Scientific Reports.

Read the publication here and download the full case study here:

Icon N1 SURFE²R Technology Customer Case Study (2020) (1.3 MB)
Nature Scientific Reports: Characterization of the Na⁺/H⁺ exchanger NhaB on the SURFE²R N1

preview the file here for download

Back

Prof. Dr. Randy Stockbridge - Interview about the SURFE²R Technology

Icon N1 “We wanted to characterize the electrophysiological characteristics of our new family of transporters to find out properties such as K_m and V_max of the substrate, and ideally to screen a set of substrates for transporter activity. However, given our difficulties in developing a transport assay, I was pessimistic that we were going to be able to move beyond radiolabeled substrates.
Contrary to my expectations, electrophysiology with the SURFE²R N1 platform worked right away. We saw very robust currents in our pilot experiments during the initial training session on the instrument.“

Prof. Dr. Randy Stockbridge, Assistant Professor
University of Michigan, USA

Download the full case study here:

Icon N1 SURFE²R Technology Customer Case Study (2019) (1.8 MB)
Replacing Radiolabeling Techniques with the SURFE²R N1

preview the file here for download

Back

データ＆アプリケーション

NCX1 - High Throughput Screening on the SURFE2R 96SE

Icon N1 SURFE²R 96SE data and applications:
Cells were kindly provided by Charles River.

NCX1 over-expressed in HEK cells was recorded on the SURFE²R 96SE. A membrane preparation of HEK cells expressing NCX1 was used on the SURFE²R 96SE. An activating buffer containing 100 μM Ca²⁺ was used to activate NCX1 (black and dark blue traces) after a negative control (0 Ca²⁺; light blue trace). NCX1 was activated three times after which a pharmacological experiment can be started to test for inhibitors. Wells are color-coded so that the user can get an overview of the succes of the experiment at a glance. Green wells depict resistance >500 MΩ, blue depicts wells have a resistance between 100 and 500 MΩ.

EAAT3 - Screening of 6 inhibitors

Icon 96SE SURFE²R 96SE data and applications:

Screening of 6 inhibitors for EAAT3. After negative control and full activation of the transporter, different concentrations of inhibitor were used. At the end the full activation was repeated.

Organic Cation Transporter 2 (OCT2) - Application of Choline, TEA and Methformin

Icon 96SE SURFE²R 96SE data and applications:

OCT2 transient currents are shown before and after the application of Choline, TEA and Methformin. A concentration-dependent activation of OCT2 could be obtained by all of the three compounds. Membranes without OCT2 (control membrane) were used as a negative control.

PepT1 - Pharmacology of Losartan

SURFE²R N96 (a predecessor model of SURFE²R S96) data and applications:

Transport by PepT1 is inhibited by the kompetitive antagonist Losartan. When activated with 20 mM Gly-Gly, the IC₅₀ of Losartan was 80.4 µM. Data were analysed with DataControl - the Analysis software tool for recordings on the SURFE²R N96 and SyncroPatch 96/ 384PE.

PepT1 - Evoked signal via Glycylglycin

SURFE²R N96 (a predecessor model of SURFE²R 96SE) data and applications:

Graphical user interface of screening software used on the SURFE²R N96. Traces of PepT1 recorded from membrane preparations of PepT1 overexpressing HEK293 cells are presented. The PepT1 signal is evoked by 20mM glycylglycin.

ウェビナー＆動画

2019 - SURFE²R as a service - closing the gap between technology and application

Icon 96SE SURFE²R 96SE Oral Presentation

Presenter:
Dr. Ralf Schwandner, CEO of Assay.Works GmbH, Germany
Assay.Works offers assay and screening services using the SURFE²R 96SE. Find out more here.

2018 - Electrophysiological Characterization of Sugar Transporters using SSM-based Electrophysiology

Icon 96SE SURFE²R 96SE and Icon N1 SURFE²R N1 Oral Presentation

Presenter:
Dr. Andre Bazzone, Application Scientist, Nanion Technologies GmbH, Germany
Andre is an expert in the field of SSM-based electrophysiology: He made his PhD at the Max-Planck-Institute of Biophysics in Frankfurt, Germany on the electrophysiological characterization of sugar transporters using the SSM-based electrophysiology in 2016. Right afterwards, he started as Application Scientist at Nanion Technologies and today he is an important member of the Nanion SURFE²R team.

2018 - Pharmacological characterization of an amino acid transporter and his bacterial homologue – a case study using the SURFE²R 96SE technology

Icon 96SE SURFE²R 96SE Oral Presentation Video

Presenter:
Dr. Thomas Licher, Head of in vitro Biology & High-Throughput Chemistry, Sanofi, IDD Frankfurt, Germany
Thomas made his PhD at the Center for Molecular Neurobiology in the group of Prof. Dr. Pongs on an HTS-technology for ion channel-modulating substances in 2002. He started to work for Sanofi-Aventis, later Sanofi as Post-doc, lab head and group leader in the field for Cardiac Safety and Lead Identification Technologies. Today, Thomas is Head of in vitro Biology & High-Throughput Chemistry at Sanofi in Frankfurt, Germany.