Simultaneous Analyte and Nucleic Acid Detection in One Lysate
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for detecting proteins and nucleic acids in cell lysates face challenges in comparing their relative amounts due to separate sample preparation and different detection techniques, making correlation difficult.
Innovation Solution
A method involving lysing cells in a multifunctional lysis buffer, followed by a proximity detection assay for analytes and a quantitative nucleic acid detection assay in the same vessel, using proximity detection probes and ligases to form ligated or hybridized probe sets for simultaneous detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If separate sample preparation methods are used for proteins and nucleic acids, then each detection method can be optimized independently, but the relative amounts of detected proteins and nucleic acids become difficult to compare
Solution Approach 1:
The patent combines separate protein and nucleic acid detection procedures into a unified workflow where both analyte types are detected in the same reaction vessel using a common lysis buffer, enabling direct comparison of relative amounts while maintaining detection accuracy for each analyte type
Solution Approach 2:
The patent develops a universal lysis buffer composition that simultaneously supports both protein stability for proximity detection assays and nucleic acid integrity for quantitative detection assays, allowing one buffer system to perform multiple functions that previously required separate specialized buffers
2Reliability
If different detection methods are used for proteins and nucleic acids, then each analyte type can be detected with specialized techniques, but correlating their relative amounts becomes difficult
Solution Approach 1:
The patent merges multiple detection systems into a single integrated assay platform where proximity detection probes for proteins and quantitative detection probes for nucleic acids coexist in the same reaction vessel, reducing the number of separate detection systems needed while maintaining reliability through specialized probe designs
3Measurement precision
If separate sample preparation is performed for proteins and nucleic acids, then each analyte can be prepared with optimal conditions, but the process requires multiple separate procedures
Solution Approach 1:
The patent combines multiple sequential sample preparation steps into a single lysis step using a multifunctional buffer, eliminating the need for separate preparation procedures for proteins and nucleic acids, thereby increasing throughput while maintaining precision through buffer composition optimized for both analyte types
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
Enables simultaneous and accurate detection of both analytes and nucleic acids in the same lysate, overcoming the limitations of separate sample preparation and detection methods.
Implementation Method 1
detecting at least one target analyte in the cell lysate using a proximity detection assay
Implementation Method 2
incubating the cell lysate with at least one ligase, such that at least one ligated proximity detection probe set is formed
Implementation Method 3
each proximity detection probe comprises at least one analyte binding moiety and at least one oligonucleotide moiety
Data Source
AI summary
Methods of detecting at least one analyte and at least one nucleic acid in a sample are provided. Reagents for carrying out the methods are also provided.