scholarly journals Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

10.3791/59886 ◽  
2019 ◽  
Author(s):  
Fernanda Festa ◽  
Joshua Labaer
Keyword(s):  
Kinase Inhibitor ◽  
Protein Microarrays ◽  
Human Protein ◽  
Inhibitor Screening ◽  
Self Assembled

scholarly journals Copper-catalyzed azide-alkyne cycloaddition (click chemistry)-based Detection of Global Pathogen-host AMPylation on Self-assembled Human Protein Microarrays

2014 ◽  
Vol 13 (11) ◽  
pp. 3164-3176 ◽  
Author(s):  
Xiaobo Yu ◽  
Andrew R. Woolery ◽  
Phi Luong ◽  
Yi Heng Hao ◽  
Markus Grammel ◽  
...  
Keyword(s):  
Click Chemistry ◽  
Protein Microarrays ◽  
Human Protein ◽  
Self Assembled

On-Chip Peptide Mass Spectrometry Imaging for Protein Kinase Inhibitor Screening

2016 ◽  
Vol 89 (1) ◽  
pp. 799-806 ◽  
Author(s):  
Young-Lai Cho ◽  
Young-Pil Kim ◽  
Jin Gyeong Son ◽  
Miyoung Son ◽  
Tae Geol Lee
Keyword(s):  
Mass Spectrometry ◽  
Protein Kinase ◽  
Mass Spectrometry Imaging ◽  
Kinase Inhibitor ◽  
Protein Kinase Inhibitor ◽  
Inhibitor Screening ◽  
Peptide Mass ◽  
On Chip ◽  
Peptide Mass Spectrometry

scholarly journals Identification of lead anti-human cytomegalovirus compounds targeting MAP4K4 via machine learning analysis of kinase inhibitor screening data

PLoS ONE ◽  
2018 ◽  
Vol 13 (7) ◽  
pp. e0201321 ◽  
Author(s):  
Blair L. Strang ◽  
Christopher R. M. Asquith ◽  
Hanan F. Moshrif ◽  
Catherine M-K Ho ◽  
William J. Zuercher ◽  
...  
Keyword(s):  
Machine Learning ◽  
Human Cytomegalovirus ◽  
Kinase Inhibitor ◽  
Inhibitor Screening ◽  
Learning Analysis

Single-step kinase inhibitor screening using a peptide-modified gold nanoparticle platform

2011 ◽  
Vol 47 (8) ◽  
pp. 2249 ◽  
Author(s):  
Shalini Gupta ◽  
Heiko Andresen ◽  
Molly M. Stevens
Keyword(s):  
Gold Nanoparticle ◽  
Kinase Inhibitor ◽  
Single Step ◽  
Inhibitor Screening

Advances in the development of human protein microarrays

2017 ◽  
Vol 14 (7) ◽  
pp. 627-641 ◽  
Author(s):  
Jessica G. Duarte ◽  
Jonathan M. Blackburn
Keyword(s):  
Protein Microarrays ◽  
Human Protein

Abstract 723: Anticancer ERK kinase inhibitor screening with molecular imaging.

2013 ◽  
Author(s):  
Lanlan Zhou ◽  
Shengliang Zhang ◽  
David T. Dicker ◽  
Alnawaz Rehemtulla ◽  
Wafik S. El-Deiry
Keyword(s):  
Molecular Imaging ◽  
Kinase Inhibitor ◽  
Inhibitor Screening ◽  
Erk Kinase

Global Assessment of B Cell Alloimmunity Using 5000 Human Protein Microarrays.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2897-2897
Author(s):  
Persis Wadia ◽  
Denong Wang ◽  
Lauren Weintraub ◽  
David B. Miklos
Keyword(s):  
Global Assessment ◽  
Chronic Gvhd ◽  
Protein Microarrays ◽  
Antibody Responses ◽  
Human Protein ◽  
Microarray Technology ◽  
Z Score ◽  
Protein Targets ◽  
Human Proteins ◽  
One Year

Abstract Allogeneic Hematopoietic Cell Transplantation (HCT) can cure hematologic malignancies through beneficial graft-v-leukemia (GVL) allo-immune responses, but its full potential is limited by graft-versus-host disease (GVHD). Recent studies demonstrate allogeneic antibodies develop against mHA encoded on the Y-chromosome, called H-Y antigens develop after sex-mismatch HCT in association with chronic GVHD and persistent disease remission. We hypothesize novel mHA can be serologically identified as targets of allogeneic antibody responses that develop after transplant and were absent pretransplant. Over 70,000 nonsynonymous single nucleotide polymorphisms (nsSNPs) encode polymorphic amino acid residues in human proteins that are potential mHA, and their analysis requires a novel high-throughput approach. ProtoArray™ (Invitrogen) displays five thousand full-length human proteins expressed as N-terminal GST-fusion proteins in a baculovirus system that are affinity purified under native conditions maintaining their cellular enzymatic activities/native conformations. These human antigens are printed in duplicate on nitrocellulose-coated glass slides. In order to determine if targets of allogeneic antibodies can be detected using microarray technology we tested three cGVHD patients for de novo Ab development after HCT using ProtoArrays™. Plasma from three male patients with AML was collected 1) prior to myeloablative HCT 2) from each HLA-identical sibling donor, and 3) one year after HCT. All three had developed extensive chronic GVHD. Plasma was diluted 1:150 and incubated on two microarrays providing replicate results. After washing, each microarray was incubated with anti-human IgG conjugated to Alexa Fluor 647 dye and detected. Negative controls include buffer, BSA, and GST, and their maximum fluorescent signals with these samples were 5,631 units with mean SD of 546. In contrast, influenza A protein is a positive control with fluorescent signal ranging 30,000–45,000 units. Correlation coefficients (R2 values) between duplicate slides ranged from 0.85 to 0.91. Data analysis was performed using Invitrogen’s Prospector Analyzer Software. Fluorescent intensity is measured for each “spot” and individual antigen results are reported as both flourescent signal intensity and a Z-score which is a measure of the intensity of a given signal relative to all of the other human protein targets reported in units of standard deviation. In order to identify human protein targets of allogeneic antibodies, each target antigen’s pretransplant Z-score was subtracted from his respective one year post-transplant Z-score yielding the “ΔZ” for that antigen. Using a conservative ΔZ of 0.1, these three patients developed new antibody responses for 67, 66, and 74 human proteins. Ninety-two percent of these proteins have known nsSNPs. No single protein was recognized by “new” antibodies in all three proteins, however polymorphic proteins Growth Arrest Specific-7 (GAS7), laminin A/C, and ribosomal protein S19 (RPS19) were recognized by two of the three patients after HCT. Results from plasma samples collected 3 and 6 months after HCT demonstrate the progression of alloimmune immune response with few new antibody responses at 3 months. Conclusion: Protein microarrays are an innovative, powerful tool for high-throughput global assessment of B cell alloimmunity after HCT. Microarray technology provides sufficient reproducibility for candidate mHA discovery. These novel mHA require validation by large clinically characterized patient samples.

Treatment with TKIs Overcomes Imatinib Resistance through the PLCgamma-1 Signaling Pathway In Imatinib Resistant Human CML Cell Lines

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4468-4468
Author(s):  
Alessandra Romano ◽  
Paolo Vigneri ◽  
Chiara Romano ◽  
Calogero Vetro ◽  
Stefania Stella ◽  
...  
Keyword(s):  
Cell Line ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Kinase Inhibitor ◽  
Conflicts Of Interest ◽  
Protein Microarrays ◽  
Resistant Cell ◽  
Resistant Cell Line ◽  
Imatinib Resistance ◽  
History Of

Abstract Abstract 4468 Suppression of BCR-ABL1 catalytic activity by the tyrosine kinase inhibitor (TKI) Imatinib Mesylate (IM) has dramatically improved the natural history of Chronic Myeloid Leukemia (CML) and - to date - represents the first and most successful example of effective anti-cancer targeted therapy {Druker, 2009}. Despite the success, patients can become resistant. Since IM-resistance in CML patients occurs despite adequate suppression of BCR-ABL activity, it is likely due to the activation of other pathways, and for this reason we need to discover novel Bcr-Abl independent pathways than can become the targets of resistant cells. Therefore we used immortalized CML human cell lines K562 and LAM84, both sensitive and resistant to Imatinib, to study the signaling in sensitive/resistant cell lines following treatment with 4 different compounds. Reverse-Phase protein microarrays were used to quantitatively map 35 cell signaling pathway endpoints, including survival, proliferation, drug resistance, apoptosis, and autophagy. Cells were incubated with IM 1uM, Dasatinib 1uM, Nilotinib 1uM or LY-29400210 uM and after 2 or 12 hours were placed in a preservative that suppresses fluctuations in kinase pathway proteins (Espina, 2008). 5/35 protein endpoints were linked together and suppressed by Dasatinib, even in the resistant cell line: PLC-y-1-Tyr783, and its upstream (ShCTyr317 SrcTyr416) and downstream targets (mTORSer2448, STAT5Tyr694, ERKThr202/Tyr204) without interfering with AKT activation status on Ser473 compared to Imatinib (p=0.0031 for K562, p= 0.042 for LAM84), Nilotinib (p=0.0034 for K562, p= 0.043 for LAM84), LY-294002 (p=0.0009 for K562, p= 0.015 for LAM84). In Imatinib-sensitive cell line LAM84 there were no differences between Dasatinib and Imatinib in the modulation of the pathway, compared to IM-sensitive K562 cell line (p=0.005), thus confirming the different profile among these two CML models. Dasatinib showed a greater suppression of the PLC-y-1 pathway compared to LY-294002 in both resistant cell lines (K562, p=0.0009 and LAM84, p=0.015). These data confirm and extend the conclusions of Markova et al. (Oncogene 2010) showing PLC-y-1 as a mechanism of death in sensitive cells. Our data showed the mechanism by which second generation TKIs (dasatinib) can overcome Imatinib resistance by suppressing the PLC-y-1 pathway. Disclosures: No relevant conflicts of interest to declare.

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