scholarly journals In Situ Target Engagement Studies in Adherent Cells

2018 ◽  
Vol 13 (4) ◽  
pp. 942-950 ◽  
Author(s):  
Hanna Axelsson ◽  
Helena Almqvist ◽  
Magdalena Otrocka ◽  
Michaela Vallin ◽  
Sara Lundqvist ◽  
...  
Keyword(s):  
Adherent Cells ◽  
Target Engagement

Monitoring Drug-Target Interactions through Target Engagement-Mediated Amplification on Arrays and in situ

2021 ◽  
Author(s):  
Rasel Al-Amin ◽  
Lars Johansson ◽  
Eldar Abdurakhmanov ◽  
Nils Landegren ◽  
Liza Löf ◽  
...  
Keyword(s):  
Drug Target ◽  
Kinase Inhibitors ◽  
Expression Profiles ◽  
Clinical Samples ◽  
Target Engagement ◽  
Rolling Circle Replication ◽  
Target Proteins ◽  
Dna Conjugates

Abstract Drugs are designed to bind their target proteins in physiologically relevant tissues and organs to modulate biological functions and elicit desirable clinical outcomes. Information about target engagement at cellular and subcellular resolution is therefore critical for guiding compound optimization in drug discovery, and for probing resistance mechanisms to targeted therapies in clinical samples. We describe a target engagement-mediated amplification (TEMA) technology, where oligonucleotide-conjugated drugs are used to visualize and measure target engagement in situ, amplified via rolling-circle replication of circularized oligonucleotide probes. We illustrate the TEMA technique using dasatinib and gefitinib, two kinase inhibitors with distinct selectivity profiles. In vitro binding by dasatinib probe to arrays of displayed proteins accurately reproduced known selectivity profiles, while their differential binding to a panel of fixed adherent cells agreed with expectations from expression profiles of the cells. These findings were corroborated by competition experiments using kinase inhibitors with overlapping and non-overlapping target specificities, and translated to pathology tissue sections. We also introduce a proximity ligation variant of TEMA in which these drug-DNA conjugates are combined with antibody-DNA conjugates to selectively investigate binding to specific target proteins of interest. This form of the assay serves to improve resolution of binding to on- and off-target proteins. In conclusion, TEMA has the potential to aid in drug development and clinical routine by conferring valuable insights in drug-target interactions at spatial resolution in protein arrays, cells and tissues.

scholarly journals Improved Procedure for Electroporation of Peptides into Adherent Cells In Situ

BioTechniques ◽  
1997 ◽  
Vol 23 (4) ◽  
pp. 644-646 ◽  
Author(s):  
Kevin L. Firth ◽  
Heather L. Brownell ◽  
Leda Raptis
Keyword(s):  
Adherent Cells

A microfluidic platform utilizing anchored water-in-oil-in-water double emulsions to create a niche for analyzing single non-adherent cells

Lab on a Chip ◽  
2019 ◽  
Vol 19 (3) ◽  
pp. 422-431 ◽  
Author(s):  
Bo Cai ◽  
Tian-Tian Ji ◽  
Ning Wang ◽  
Xin-Bo Li ◽  
Rong-Xiang He ◽  
...  
Keyword(s):  
Adherent Cells ◽  
Microfluidic Platform ◽  
Double Emulsions ◽  
Oil In Water ◽  
On Chip

Water-in-oil-in-water double emulsions (W/O/W DEs) are generated to encapsulate non-adherent cells and anchored in an array on-chip for in situ assays.

scholarly journals Recovery of Adherent Cells after In Situ Electroporation Monitored Electrically

BioTechniques ◽  
2002 ◽  
Vol 33 (2) ◽  
pp. 348-357 ◽  
Author(s):  
Joachim Wegener ◽  
Charles R. Keese ◽  
Ivar Giaever
Keyword(s):  
Adherent Cells ◽  
In Situ Electroporation

Impedance analysis of adherent cells after in situ electroporation: Non-invasive monitoring during intracellular manipulations

2011 ◽  
Vol 26 (12) ◽  
pp. 4720-4727 ◽  
Author(s):  
Judith A. Stolwijk ◽  
Christoph Hartmann ◽  
Poonam Balani ◽  
Silke Albermann ◽  
Charles R. Keese ◽  
...  
Keyword(s):  
Impedance Analysis ◽  
Adherent Cells ◽  
Invasive Monitoring ◽  
Non Invasive ◽  
In Situ Electroporation

IN SITU LABELING OF ADHERENT CELLS WITH PKH26

2000 ◽  
Vol 36 (1) ◽  
pp. 4 ◽  
Author(s):  
GYUN MIN LEE ◽  
STEVE FONG ◽  
KARL FRANCIS ◽  
DUK JAE OH ◽  
BERNHARD O. PALSSON
Keyword(s):  
Adherent Cells

scholarly journals Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells

2015 ◽  
Vol 10 (6) ◽  
pp. 554-562 ◽  
Author(s):  
Lindsey Hanson ◽  
Wenting Zhao ◽  
Hsin-Ya Lou ◽  
Ziliang Carter Lin ◽  
Seok Woo Lee ◽  
...  
Keyword(s):  
Adherent Cells ◽  
Nuclear Mechanics

Electroporation of Adherent Cells In Situ for the Introduction of Nonpermeant Molecules

2003 ◽  
pp. 93-114 ◽  
Author(s):  
Leda H. Raptis ◽  
Kevin L. Firth ◽  
Heather L. Brownell ◽  
Andrea Todd ◽  
W. Craig Simon ◽  
...  
Keyword(s):  
Adherent Cells

Design and Implementation of a Microelectrode Assembly for Use on Noncontact In Situ Electroporation of Adherent Cells

2012 ◽  
Vol 245 (10) ◽  
pp. 617-624 ◽  
Author(s):  
Tomás García-Sánchez ◽  
Beatriz Sánchez-Ortiz ◽  
Ingrid Vila ◽  
Maria Guitart ◽  
Javier Rosell ◽  
...  
Keyword(s):  
Adherent Cells ◽  
Design And Implementation ◽  
In Situ Electroporation
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