Name: rapid homogeneous detection of biological assays using magnetic modulation biosensing system
Uploaded: 2010-06-13T17:00:00
Description: Watch this Scientific Journal Video about Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System at JoVE.com

<p class="jove_content">This work was partly supported by the Ishaya Horowitz Fund.</p>

<p class="jove_title">IL-8 assay:</p>
<ol>
  <li>The reaction mixture included the following four components in 100 &mu;l final volume of assay buffer in their respective concentrations. 10 &mu;l of magnetic beads with capture antibody at 100 beads/&mu;l final concentrations, 2 &mu;l of biotinylated IL-8 antibody at 1 nano-gram/&mu;l final concentration, 1 &mu;l of streptavidin fluorescent protein at 20 nano-gram/&mu;l final concentration, and 1 &mu;l of IL-8 target at 0.48 pico-gram/&mu;l. The components are added one by one to the assay buf...

<ol>
	<li>Danielli, A., Arie, A., Porat, N., Ehrlich, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+fluorescent-labeled+probes+at+sub-picomolar+concentrations+by+magnetic+modulation.">Detection of fluorescent-labeled probes at sub-picomolar concentrations by magnetic modulation.</a> <em style='font-style: italic'>Optics Express</em>. <b>16</b>, 19253-19259 (2008).</li><li>Danielli, A., Porat, N., Arie, A., Ehrlich, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+homogenous+detection+of+the+Ibaraki+virus+NS3+cDNA+at+pico-molar+concentrations+by+magnetic+modulation.">Rapid homogenous detection of the Ibaraki virus NS3 cDNA at pico-molar concentrations by magnetic modulation.</a> <em style='font-style: italic'>Biosensors and Bioelectronics</em>. <b>25</b>, 858-863 (2009).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Bio-Rad "Human angiogenesis assay panel"</em>. , .</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Instruction+Manual.">Instruction Manual.</a> <em style='font-style: italic'>Bio-Plex Precision Pro Cytokine Assay</em>. , .</li></ol>

<p class="jove_content">In summary, we showed that the MMB system can be used to detect the presence of IL-8 target at low concentrations (0.5 pico-gram is the detection limit of the Bio-Plex Precision Pro cytokine assays [3,4]). The ability of the system is not limited to IL-8 and can be used to detect other proteins. The advantages of the MMB system are the ability to detect the target rapidly and without any separation or washing step. Thus, it facilitates the detection process and allows the system to be used in field applications.</p>

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Bio-Plex Pro Human Cytokine IL-8 set</td>
<td>&nbsp;</td>
<td>Bio-Rad</td>
<td>171-B5008M</td>
<td>&nbsp;</td>
<tr>
<td>Recombinant human  IL-8 target</td>
<td>&nbsp;</td>
<td>Thermo Scientific</td>
<td>R202625</td>
<td>&nbsp;</td>
<tr>
<td>Biotinylated monoclonal mouse anti human IL-8 antibody G265-8</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>554718</td>
<td>&nbsp;</td>
<tr>
<td>Streptavidin coupled fluorescent protein</td>
<td>&nbsp;</td>
<td>Bio-Rad</td>
<td>Streptavidin PE</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
	

rapid homogeneous detection of biological assays using magnetic modulation biosensing system

A magnetic modulation biosensing system (MMB) [1,2] rapidly and homogeneously detected biological targets at low concentrations without any washing or separation step. When the IL-8 target was present, a 'sandwich'-based assay attached magnetic beads with IL-8 capture antibody to streptavidin coupled fluorescent protein via the IL-8 target and a biotinylated IL-8 antibody. The magnetic beads are maneuvered into oscillatory motion by applying an alternating magnetic field gradient through two electromagnetic poles. The fluorescent proteins, which are attached to the magnetic beads are condensed into the detection area and their movement in and out of an orthogonal laser beam produces a periodic fluorescent signal that is demodulated using synchronous detection. The magnetic modulation biosensing system was previously used to detect the coding sequences of the non-structural Ibaraki virus protein 3 (NS3) complementary DNA (cDNA) [2]. The techniques that are demonstrated in this work for external manipulation and condensation of particles may be used for other applications, e.g. delivery of magnetically-coupled drugs <em>in-vivo</em> or enhancing the contrast for <em>in-vivo</em> imaging applications.

Watch this Scientific Journal Video about Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System at JoVE.com

Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System

<p class="jove_content">Magnetic modulation biosensing system is utilized to rapidly, sensitively and simply detect biological assays, such as DNA molecules and proteins.</p>

A magnetic modulation biosensing system (MMB) [1,2] rapidly and homogeneously detected biological targets at low concentrations without any washing or ...

Research

JoVE Journal

Biology

ELIME (Enzyme Linked Immuno Magnetic Electrochemical) Method for Mycotoxin Detection

ELIME Enzyme Linked Immuno Magnetic Electrochemical Method for Mycotoxin Detection

Immunoassays are a valid alternative to the more expensive and time consuming quantitative HPLC or GC1, 2 methods for the screening detection of hazardous ...

Analysis of Schwann-astrocyte Interactions Using <em>In Vitro</em> Assays

Analysis of Schwann-astrocyte Interactions Using In Vitro Assays

Schwann cells are one of the commonly used cells in repair strategies following spinal cord injuries. Schwann cells are capable of supporting axonal ...

High-throughput Screening and Biosensing with Fluorescent <em>C. elegans</em> Strains

High-throughput Screening and Biosensing with Fluorescent C. elegans Strains

High-throughput screening (HTS) is a powerful approach for identifying chemical modulators of biological processes.  However, many compounds identified in ...

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation BiFC System

We have developed a BiFC technique to test the interaction between two proteins in vivo. This is accomplished by splitting a yellow fluorescent protein ...

Rapid Protocol for Preparation of Electrocompetent <em>Escherichia coli</em> and <em>Vibrio cholerae</em>

Rapid Protocol for Preparation of Electrocompetent Escherichia coli and Vibrio cholerae

Electroporation has become a widely used method for rapidly and efficiently introducing foreign DNA into a wide range of cells. Electrotransformation has ...

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Zebrafish is a powerful vertebrate model system for studying development, modeling disease, and performing drug screening. Recently a variety of genetic ...

Methods for Skin Wounding and Assays for Wound Responses in <em>C. elegans</em>

Methods for Skin Wounding and Assays for Wound Responses in C. elegans

The C. elegans epidermis and cuticle form a simple yet sophisticated skin layer that can repair localized damage resulting from wounding. Studies of wound ...

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Digital assays are powerful methods that enable detection of rare cells and counting of individual nucleic acid molecules. However, digital assays are ...

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC)

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents HPHC

Cigarette smoke (CS) is a major risk factor for cardiovascular and lung diseases. Because CS is a complex aerosol containing more than 7,000 chemicals1 it ...