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MANUAL PATCH CLAMP

The gold standard, in quality, for assessing interactions between compounds and ion channels is considered the manual (i.e., conventional) patch clamp electrophysiology. 

The IC50 concentration-response assay provides important benchmarks for structure-activity evaluation, whereas a Manual Patch Screen tests compounds within structural or therapeutic groups against one or more ion channels. 

To find the concentration dose dependent, our team performs the manual patch clamp onto cells with homologous or heterologous expression of different ion channels for assessment of compound's interactions with the channel in continuous perfusion. 

Manual patch-clamp equipped with Multiclamp 700A amplifier (Molecular Devices, Sunnyvale, CA), Digidata 1322A and pClamp 8.2 software (Molecular Devices, Sunnyvale, CA).

Assay features

Number of concentration

4-6

Number of replicates per concentration

≥ 5

Vehicle control (max concentration n ≥ 5)

optional

Positive control (n ≥ 5)

included

Families of available ion channels

K, Na, Ca

The recordings can be performed at either physiological or room temperature.


References:

1.Gasparoli L, D'Amico M, Masselli M, Pillozzi S, Caves R, Khuwaileh R, Tiedke W, Mugridge K, Pratesi A, Mitcheson JS, Basso G, Becchetti A, Arcangeli A. New pyrimido-indole compound CD-160130 preferentially inhibits the KV11.1B isoform and produces antileukemic effects without cardiotoxicity. Mol Pharmacol. 2015 Feb;87(2):183-96

2.Pillozzi S, Gasparoli L, Stefanini M, Ristori M, D'Amico M, Alessio E, Scaletti F, Becchetti A, Arcangeli A, Messori L. NAMI-A is highly cytotoxic toward leukaemia cell lines: evidence of inhibition of KCa 3.1 channels. Dalton Trans. 2014 Aug 28;43(32):12150-5

3.Crociani O, Zanieri F, Pillozzi S, Lastraioli E, Stefanini M, Fiore A, Fortunato A, D'Amico M, Masselli M, De Lorenzo E, Gasparoli L, Chiu M, Bussolati O, Becchetti A, Arcangeli A. hERG1 channels modulate integrin signaling to trigger angiogenesis and tumor progression in colorectal cancer. Sci Rep. 2013 Nov 25;3:3308.

4.Biagiotti T, D'Amico M, Marzi I, Di Gennaro P, Arcangeli A, Wanke E, Olivotto M. Cell renewing in neuroblastoma: electrophysiological and immunocytochemical characterization of stem cells and derivatives. Stem Cells. 2006, 24(2), 443-53

5.D’Amico M, Biagiotti T, Fontana L, Restano-Cassulini R, Lasagna N, Arcangeli A, Wanke E and Olivotto M. A HERG current sustains a cardiac-type action potential in neuroblastoma S cells. Biochem. Biophys. Res. Commun., 2003, 302, 101-108.