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Luminescent biosensors for real-time monitoring of intracellularcAMP.

Binkowski BF,Fan F,Wood KV.

Source

Promega Corporation, Madison, WI, USA. brock.binkowski@promega.com

Abstract

G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) comprise a diverse class of

signaling molecules involved in cellular physiology and pathology. In recent years, intracellular biosensors

have been developed to monitor changes in intracellular cAMP in real time, facilitating studies on the

mechanisms of GPCR activation and desensitization in living cells. However, methods based on

fluorescence can show limitations in response dynamics together with being difficult to perform. Here we

present the use of genetically encoded, luminescent biosensors that allow a facile, non-lytic assay format

for monitoring cAMP dynamics in living cells.

Nanowire biosensors for label-free, real-time, ultrasensitive proteindetection.

Zheng G,Lieber CM.

Source

Laboratory of Advanced Materials, Department of Chemistry, Fudan University, Shanghai, China. gfzheng@fudan.edu.cn

Abstract

Sensitive and quantitative analysis of proteins is central to disease diagnosis, drug screening, and

proteomic studies. Among recent research advances exploiting new nanomaterials for biomolecule

analysis, silicon nanowires (SiNWs), which are configured as field-effect transistors (FETs), have

emerged as one of the most promising and powerful platforms for label-free, real-time, and highly

sensitive electrical detection of proteins as well as many other biological species. Here, we describe adetailed protocol for realizing SiNW biosensors for protein detection that includes SiNW synthesis, FET

device fabrication, surface receptor functionalization, and electrical sensing measurements. Moreover,

incorporating both p-type and n-type SiNWs in the same sensor array provides a unique means of

internal control for sensing signal verification.

Near real-time, on-site, quantitative analysis of PAHs in the aqueous environment using anantibody-based biosensorKeywords:

Biosensor;

PAH;

Immunoassay;

Monoclonal antibodies;

Real-time monitoring

Abstract

Rapid, on-site, quantitative assessments of dissolved polycyclic aromatic hydrocarbons (PAHs) were demonstrated

for two field applications. The platform, a KinExA Inline Sensor (Sapidyne Instruments), employed the monoclonal

anti-PAH antibody, 7B2.3, which has specificity for 3- to 5-ring PAHs. A spatial study was conducted near a dredging

http://www.ncbi.nlm.nih.gov/pubmed?term=%22Binkowski%20BF%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Binkowski%20BF%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Fan%20F%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Fan%20F%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Fan%20F%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Wood%20KV%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Wood%20KV%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Wood%20KV%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Zheng%20G%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Zheng%20G%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Lieber%20CM%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Lieber%20CM%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Lieber%20CM%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Lieber%20CM%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Zheng%20G%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Wood%20KV%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Fan%20F%22%5BAuthor%5Dhttp://www.ncbi.nlm.nih.gov/pubmed?term=%22Binkowski%20BF%22%5BAuthor%5D

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site where contaminated sediments were being removed, and a temporal study was performed during a rainfall event.

Most importantly, the generation of near real-time data guided management decisions in the field and determined

proper sampling protocols for conventional analyses. The method was able to determine PAH concentrations as low

as 0.3g/L, within 10min of sample acquisition, and to assess 80+ samples (not including standards and blanks) in

less than 3 d. These results were compared with a laboratory-based gas chromatographymass spectrometry

method in which a wide array of PAHs, including alkylated homologs, were examined. This system shows great

promise as a field instrument for the rapid monitoring of PAH pollution. Environ. Toxicol. Chem. 2011; 30:15571563.

2011 SETAC

A cell-based biosensor for real-time detection of cardiotoxicity using lensfree imaging

Sang Bok Kim , Hojae Bae , Jae Min Cha , Sang Jun Moon , Mehmet R. Dokmeci , Donald M. Cropek and Ali

Khademhosseini

Lab Chip, 2011, 11, 1801-1807

DOI: 10.1039/C1LC20098D

Received 03 Feb 2011, Accepted 23 Mar 2011

First published on the web 11 Apr 2011

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A portable and cost-effective real-time cardiotoxicity biosensor was developed using a CMOS imaging module

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evaluate the engineered system, the effects of two different drugs, isoprenaline and doxorubicin, on the beating rate

and beat-to-beat variations of ESC-derived cardiomyocytes were measured. The detection system was used to

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the beating rates increased upon the addition ofisoprenaline but decreased for cultures supplemented

with doxorubicin. Moreover, the response time for both the beating rates and the beat-to-beat variability of ESC-

derivedcardiomyocytes under treatment of isoprenaline was shorter than for doxorubicin, although the amount

of isoprenaline used in the measurement was three orders of magnitude lower than that of doxorubicin. Given its

ability to perform real-time cell monitoring in a simple and inexpensive manner, the proposed system may be useful

for a range of cell-based biosensing applications.

Webcam measures medications effects on heartCompiled by BioPhotonics staff

BOSTON A tool using basic webcam technology enables real-time study of medications effects onheart cells called cardiomyocytes including any drug-induced changes in the beating rate. Thetechnology could help drug makers and prescribers address a side effect of drugs called cardiotoxicity an unhealthy change in the way the heart beats.

Scientists at Brigham and Womens Hospital (BWH) have developed a cost-effective, portable cell-based biosensor for real-time cardiotoxicity detection using an image sensor from a basic webcam.Using the biosensor to monitor the beating rates and to detect any drug-induced changes, theyintroduced various drugs to cardiomyocytes derived from mouse stem cells.

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Using an image sensor from a basic webcam, scientists at Brigham and Womens Hospital havedeveloped a portable cell-based biosensor for real-time detection of cardiotoxicity.

The new technique provides a simple approach to performing evaluative studies of different drugseffects on cardiac cells.

Assessing the toxic effects of new drugs during the early phases of drug development can acceleratethe drug discovery process, resulting in significant cost and time savings, and leading to fastertreatment discovery, said Ali Khadem-hosseini, PhD, of the Center for Biomedical Engineering at theDepartment of Medicine at BWH.

In addition, the technology could play a role in personalized medicine. The biosensor could be used tomonitor cardiac cells that have been introduced to medication, providing a glimpse of how the drugaffects the individuals heart, which could help doctors shape the treatment plan for that patient.

The researchers hope to combine the detection system with their microwell arrays to screen thousandsof drugs simultaneously in a fast, reliable manner, Khademhosseini said.

ARTICLE DISCUSSION