High-throughput Cell-free Assay for Detection of Quorum Sensing Signals, Acyl-Homoserine Lactones (AHLs)

Description:

Reference #:  00653

The University of South Carolina is offering licensing opportunities for high-throughput cell-free assay for detection of QS chemical signals.

Background/Challenges:

Quorum sensing (QS) is an emerging research field in health and environmental sciences during the past decade. QS is the ability of bacteria to communicate and coordinate behavior such as biofilm formation via signaling molecules called auto-inducers (e.g. acyl-homoserine lactones (AHLs)). Therefore, it is conceivable that quorum sensing inhibition represent a new, novel strategy to disrupt biofilm formation to combat many diseases.

A simple, sensitive and rapid assay for the detection of AHLs is urgently needed in quorum sensing research. There are many methods developed to detect AHLs, such as whole cell assays using biosensors, thin-layer chromatography (TLC), gas chromatography (GC), high performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS). Among those methods, whole cell bioassay using Agrobacterium tumefaciens NTL4 has been the most popular sensitive method because this method doesn't require expensive instruments such as HPLC and LC/MS and a skilled scientist. However, there are disadvantages of the whole cell bioassay:

1)    It requires labor intensive cell conditioning

2)    It takes at least 24hours for detection of AHLs

3)    It may be difficult to screen the inhibitors of QS when the test substance inhibits cell growth itself and

4)    It is not accurately quantitative but qualitative

Invention Description:

The subject invention is a high-throughput cell-free assay for detection of QS chemical signals, AHLs, using Agrobacterium tumefaciens NTL4. With this system, it is possible to screen many samples for the presence of AHLs using microtiter plates by simple addition of Agrobacterium tumfaciens NTL4 cell-free extract solution. AHLs can be detected using the micro-plate reader within 3 hours. This assay system eliminates time consuming cell conditioning before each day. Also, cell-free extract can be stored in a -20oC freezer for a long period of time.

Advantages and Benefits:

The cell-free assay system for detecting quorum sensing signals eliminates time-consuming bio-sensor cell conditioning required before a whole cell bioassay. It therefore simplifies the assay procedure and saves time. This protocol is very suitable to screen not only quorum sensing colony and quorom sensing inhibitors but also AHLs receptor proteins and co-factors in a large number of samples using a 96 well plate in quorum sensing research. By using the fluorogenic substrate instead of X-gal, the system can increase its sensitivity to AHLs. This method is simple, cost effective, and can easily be applied to detect the presence of AHLs in the environmental samples in the filed condition.

Potential Applications:

The invention can find application in research where the cell-free assay is used for the detection of Quorum sensing (QS) chemical signals. Since this method uses the cell-free system, it can also be utilized as a great educational tool for students to understand the process of quorum sensing.

 

Patent Information:
Title App Type Country Serial No. Patent No. File Date Issued Date Expire Date Patent Status
High-throughput In Vitro Translation (Cell Lysate Based) Assay for Detecting Quorum Sensing Signals Utility United States 12/180,857 8,546,071 7/28/2008 10/1/2013 8/29/2030  
For Information, Contact:
Technology Commercialization
University of South Carolina
technology@sc.edu
Inventors:
Alan Decho
Tomohiro Kawaguchi
Yung-Pin Chen
Keywords:
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