Affinity-based detection of biological targets
Inventors
Kulagina, Nadezhda V • Taitt, Chris Rowe • Anderson, George P • Ligler, Frances S
Assignees
Publication Number
US-8658372-B2
Publication Date
2014-02-25
Expiration Date
2026-02-06
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Abstract
A method of biochemical identification by: providing a plurality of capture species bound to one or more substrates and suspected of having one or more biological targets affinity bound to at least one capture species; detecting which capture species contain bound biological targets to generate a binding pattern; and identifying the biological target based on the binding pattern. The capture species are independently selected from the group consisting of antimicrobial peptides, cytotoxic peptides, antibiotics, and combinations thereof. A device having the capture species bound to the substrates. At least two of the capture species are capable of multi-specific binding to one or more biological targets and may have overlapping but not identical affinity properties.
Core Innovation
The invention is a biochemical identification method and a device that uses a plurality of capture species bound to one or more substrates to detect biological targets based on their binding patterns. The capture species are independently selected from antimicrobial peptides, cytotoxic peptides, antibiotics, and their combinations. The method involves exposing these capture species to samples suspected of containing biological targets, detecting which capture species have bound targets to generate a binding pattern, and identifying the biological target using this pattern, potentially by comparing it to a database through a pattern recognition algorithm.
Traditional antibody-based detection systems suffer from issues such as temperature sensitivity, the need for specific antibodies for each target, complexity in multiplexed assays, cost and time in monoclonal antibody production, and limited stability for long-term field applications. These limitations motivate the development of a multiplexed system using antimicrobial peptides and antibiotics, which naturally exhibit semi-selective binding properties, high stability in adverse conditions, and the ability to bind multiple microbial species. The invention addresses these problems by substituting or complementing antibodies with antimicrobial peptides and antibiotics, thereby offering increased stability, reduced complexity, and the potential for multiplexed analyte screening.
The capture species used are not combinatorially derived or randomly generated but are naturally occurring or derivatives thereof, with defined secondary structures and overlapping specificities. These semi-selective properties enable the detection of a larger number of targets with fewer recognition molecules. The interaction mechanism is affinity-based, not enzymatic, and the system can detect various biological targets including bacteria, viruses, fungi, rickettsiae, and toxins. The invention can be implemented on various detection platforms, using different immobilization chemistries and detection modalities, such as fluorescence or reagent-less detection methods.
Claims Coverage
The patent includes one independent claim defining the biochemical screening method of identifying biological targets using capture species, along with several dependent claims describing additional features and embodiments.
Use of antimicrobial peptides and antibiotics as capture species
Providing a plurality of capture species independently selected from antimicrobial peptides, antibiotics, and their combinations, bound to one or more substrates for biochemical screening.
Generation and use of binding patterns
Exposing the substrates to known biological targets, allowing binding to capture species, detecting binding patterns, building a database of such patterns, and comparing unknown target binding patterns to this database via a pattern recognition algorithm.
Capture species with overlapping but not identical affinities
At least two of the capture species have overlapping but not identical affinity properties to enable pattern-based target identification.
Detection methods using labeled targets or tracer species
The biological target may have a label capable of producing an opto-electronic signal, or detection may involve tracer species comprising a recognition element and a signal generating element such as fluorophores, chromophores, fluorescent nanospheres, or enzymes.
Reagent-less detection options
The detecting step may alternatively perform reagent-less analysis methods including opto-electronics, surface plasmon resonance, interferometry, and quartz crystalline microbalance.
Use of cytotoxic peptides as capture species
Inclusion of cytotoxic peptides as additional capture species.
Broad capture species selection
Capture species selection includes a comprehensive list of natural and derivative antimicrobial peptides and antibiotics such as alamethicin, bacitracin, cecropins, magainins, polymyxins, vancomycin, and others, including naturally occurring peptides.
Target biological entities
Biological targets include bacteria, fungi, viruses, rickettsiae, toxins, and their combinations.
The independent claim covers a biochemical screening method that uses antimicrobial peptides and antibiotics as capture species bound to substrates to generate and analyze binding patterns for identifying biological targets. The features include overlapping affinity properties of capture species, diverse detection methods, and comprehensive lists of capture species and biological targets.
Stated Advantages
Improved stability and resistance to adverse environmental conditions compared to antibody-based assays.
Capability for simultaneous detection and identification of a larger number of targets with fewer recognition molecules due to semi-selective, overlapping binding properties.
Potential for equivalent or superior detection sensitivity compared to antibody-based systems.
Reduced complexity and logistical burdens in multiplexed assays by eliminating the need for specific antibodies for each target.
Suitability for use in fielded sensor applications due to enhanced stability and robustness.
Potential for therapeutic and decontamination applications through binding pattern indications of target sensitivity to antimicrobial peptides and antibiotics.
Documented Applications
Detection and identification of bacteria, fungi, viruses, rickettsiae, and toxins using array biosensors and Luminex flow cytometry systems.
Multiplexed biochemical assays for rapid detection in clinical diagnostics, environmental monitoring, and other field applications.
Use in biosensors that currently utilize antibodies for target recognition, replacing or complementing antibodies with antimicrobial peptides and antibiotics.
Direct detection of pathogens and toxins through affinity-based binding patterns.
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