Single-Molecular Homogenous Amplified Detection in Confined Volumes

Case 2014-180

 

UCLA researchers have developed a novel method of detecting molecules of interest which can be performed without washing steps or any solid-phase reaction.

 

BACKGROUND

 

The typical sandwich ELISA utilizes adsorbed antibodies on a surface that capture antigens and a secondary antibody with sequential washes to provide a detection signal. Others have used antibody-coated beads coupled with femtoliter-sized wells to fractionate the sample and generate a potentially more robust assay with a digital output. However, since these assays require one bead per well, the bead capture efficiency limits the sensitivity of the assay. Additionally, both traditional and digital ELISAs require sequential addition of reagents for binding as well as a number of wash steps to remove unbound reagents. Existing assays require detection antibodies to be immobilized on a surface, including bead-based assays.

 

INNOVATION

 

Researchers at UCLA have identified a novel method of detecting molecules of interest which can be performed without washing steps or any solid-phase reaction. Since this assay does not require a surface or beads to adsorb to and other sample preparation steps, this assay is ideally suited for point-of-care use. Instead, this assay requires specially modified antibodies to allow for amplification of the detection event. This assay does not require a number of wash steps or sequential addition of reagents, which is typical of current assays; instead this assay just requires a single mix step of the reagents with the target sample.

 

APPLICATIONS

  • Clinical diagnostics in hospitals or private labs
  • Point of care diagnostics
  • Research

 

ADVANTAGES

  • Does not require any wash steps, decreasing the amount of reagents needed and associated costs, as well as increasing ease of use, efficiency, and scalability
  • Sensitivity and range of the assay can be adjusted to optimize molecule detection
  • High sensitivity and specificity, with a low limit of detection

 

STATE OF DEVELOPMENT

 

Various aspects of the assay have been developed, including the microfluidics aspect that is important to enable the homogenous assay. Testing of the reaction components in a well plate as well as within the device has been carried out, but not optimized.

Patent Information: