Engineered Opioid Biosensors via Mutated PBP Domains
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Solution Overview
Problem
Current methods for monitoring opioid levels in humans are laborious and lack suitable opioid-binding moieties for continuous monitoring, complicating the management of opioid use disorder due to incomplete knowledge of cellular tolerance mechanisms and individual variability in responses to opioids.
Innovation Solution
Development of engineered opioid biosensors comprising mutated periplasmic binding protein domains that undergo detectable conformational changes upon opioid binding, enabling sensitive and selective detection of opioids in biological samples, including those specific to kappa, delta, or mu opioid receptors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional blood draw and LC-MS quantification methods are used for opioid monitoring, then measurement precision is achieved, but loss of time and ease of operation deteriorate due to laborious procedures and requirement for clinical setting visits
Solution Approach 1:
The patent replaces the mechanical/biochemical LC-MS quantification system with an optical detection system using fluorescent biosensors. The biosensors utilize fluorescence emission to detect opioid binding events, substituting complex mechanical separation and detection mechanisms with optical signal transduction that can be monitored in real-time without requiring sample processing or clinical setting equipment.
Solution Approach 2:
The engineered biosensors perform self-detection by undergoing conformational changes upon opioid binding that directly modulate their own fluorescent signal. The biosensors autonomously transduce binding events into measurable optical signals without requiring external processing steps, reagents, or complex instrumentation, enabling continuous monitoring in a self-sufficient manner.
2Measurement precision
If conventional blood draw and LC-MS quantification methods are used for opioid monitoring, then measurement precision is achieved, but ease of operation deteriorates due to requirement for clinical setting visits
Solution Approach 1:
The patent replaces the mechanical/biochemical LC-MS quantification system with an optical detection system using fluorescent biosensors. The biosensors utilize fluorescence emission to detect opioid binding events, substituting complex mechanical separation and detection mechanisms with optical signal transduction that can be monitored in real-time without requiring sample processing or clinical setting equipment.
Solution Approach 2:
The engineered biosensors perform self-detection by undergoing conformational changes upon opioid binding that directly modulate their own fluorescent signal. The biosensors autonomously transduce binding events into measurable optical signals without requiring external processing steps, reagents, or complex instrumentation, enabling continuous monitoring in a self-sufficient manner.
3Measurement precision
If engineered biosensors with mutated PBP domains are developed, then sensitivity and selectivity for opioid detection are improved, but device complexity increases due to protein engineering requirements
Solution Approach 1:
The patent applies parameter changes by systematically mutating specific amino acid residues in the PBP domain at defined positions to optimize opioid binding affinity and selectivity. By changing the chemical parameters (amino acid side chains) at key binding sites, the biosensor achieves enhanced sensitivity and specificity for detecting pharmacologically relevant opioid concentrations while maintaining a relatively simple overall protein structure.
Solution Approach 2:
The patent applies local quality by introducing mutations at specific localized positions within the PBP domain that are functionally equivalent to known opioid-binding residues. Rather than redesigning the entire protein, localized mutations at critical binding sites (positions functionally equivalent to K10, N11, Q15, T43, T68, T325, K330, D341, Y357, A360, E391, R395, V405, F436, H455, and D490) confer enhanced opioid selectivity while preserving the overall protein fold and function.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The biosensors provide continuous monitoring with improved sensitivity and selectivity for opioids, allowing for precise detection in pharmacologically relevant concentrations, facilitating better management of opioid use disorder and overdose prevention.
Implementation Method 1
the opioid biosensor is capable of undergoing a detectable conformational change upon binding to an opioid
Data Source
AI summary
Disclosed herein include engineered opioid biosensors, and related compositions, vectors, cells, and systems. Also disclosed include methods that provide opioid biosensors with sensitivity and selectivity suitable for continuous opioid monitoring as well as the use of the opioid biosensors for detecting one or more specific opioids. The opioid biosensors, which are capable of undergoing a detectable conformational change upon binding to an opioid, can each comprise a first periplasmic binding protein (PBP) domain and a second PBP domain connected to the first PBP domain, wherein at least one of the first PBP domain and the second PBP domain comprises one or more mutations.