Methods and compositions for backscattering interferometry
Inventors
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Assignees
Vanderbilt University • Base Pair Biotechnologies Inc
Member
Base Pair BiotechnologiesBase Pair Biotechnologies specializes in custom aptamer discovery and development for research, diagnostics, therapeutics, and industrial applications. The company leverages proprietary multiplex selection, advanced bioinformatics, and chemical modification techniques to develop high-affinity and selective nucleic acid aptamers. Base Pair enables affinity reagent development, biosensor design, and molecular detection for a broad range of targets and partners across academia and industry.
Base Pair Biotechnologies specializes in custom aptamer discovery and development for research, diagnostics, therapeutics, and industrial applications. The company leverages proprietary multiplex selection, advanced bioinformatics, and chemical modification techniques to develop high-affinity and selective nucleic acid aptamers. Base Pair enables affinity reagent development, biosensor design, and molecular detection for a broad range of targets and partners across academia and industry.
Publication Number
US-11078489-B2
Publication Date
2021-08-03
Expiration Date
Abstract
The present invention relates to methods and compositions for enhancing backscattering interferometry (BSI) in detection of biomolecular interactions, particularly to methods and compositions for enhancing BSI utilizing label-free aptamers, and more particularly to methods and compositions for enhancing BSI utilizing high conformational change aptamers, which may change in conformation when the aptamers bind to their target molecules.
Core Innovation
The present invention relates to methods and compositions for enhancing backscattering interferometry (BSI) in detection of biomolecular interactions, particularly to methods and compositions for enhancing BSI utilizing label-free aptamers, and more particularly to methods and compositions for enhancing BSI utilizing high conformational change aptamers, which may change in conformation when the aptamers bind to their target molecules. In one aspect, methods and compositions for enhancing BSI may employ label-free aptamers which may generally exhibit a large conformational change between their free, unbound state and their bound state when bound to their target molecule.
The background describes that surface plasmon resonance (SPR) measures localized change in refractive index but "is a heterogeneous method that requires complicated surface chemistry, immobilization, possible modification of one of the species being examined, and expensive gold-plated slides." The background further states that biolayer interferometry (BLI) "relies on surface immobilization and suffers from similar problems to SPR."
To address these limitations, the invention utilizes at least partially self-complementary aptamers, including nucleic acid hairpins with a self-hybridizing stem region and a non-self-hybridizing loop region, that may partially hybridize in the absence of target and dehybridize upon target binding to generate a large conformational change. The stem region may be designed to form a predetermined number of Watson-Crick hybridizing pairs and its length may be tailored to modulate sensitivity, and BSI signal may also be improved in an intermolecular system by utilizing an aptamer and a separate at least partially complementary nucleic acid.
Claims Coverage
The patent includes one independent claim with two main inventive features.
Hairpin aptamer comprising SEQ ID No. 1
a hairpin aptamer having a non-self-hybridizing loop region and self-hybridizing stem region, said hairpin aptamer comprising a non-naturally occurring sequence comprising the sequence of SEQ ID No. 1;
Stem region dehybridization elicits conformational change
wherein said self-hybridizing stem region dehybridizes upon binding of said hairpin aptamer to a target molecule to elicit a conformational change in said hairpin aptamer.
The independent claim covers a non-labeled molecular beacon defined by a hairpin aptamer comprising SEQ ID No. 1 and a self-hybridizing stem region that dehybridizes upon target binding to elicit a conformational change.
Stated Advantages
Enhancing backscattering interferometry (BSI) in detection of biomolecular interactions by utilizing label-free, high conformational change aptamers.
BSI is a label-free, free-solution technology that generally allows for a broad range of binding affinities to be measured in a matter of hours without a priori knowledge of the binding system.
Because the BSI signal is a product of inherent properties of the sample, there is generally no need for surface-immobilization or labeling to characterize a binding interaction quantitatively.
Aptamers can be designed and produced rapidly and inexpensively, are stable for long periods without the strict storage requirements of antibodies, can be produced by purely synthetic means with virtually no lot-to-lot variability, and allow facile modifications to confer nuclease stability.
Documented Applications
Detection of biomolecular interactions using backscattering interferometry (BSI).
Use of aptamers as sensors, therapeutic tools, and cellular process regulators, and to guide drugs to their specific cellular targets.
Characterization of interactions between aptamers and their target molecules and measuring binding affinity.
Exemplary aptamer systems demonstrated binding to tenofovir and to HIV capsid protein p24.
Interested in licensing this patent?