Avidin-Biotin Probe Complex Assembly for Sensitive Detection
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Solution Overview
Problem
Existing methods for producing probe-label conjugates with high sensitivity and stability are hindered by increased blank reactions and reproducibility issues due to the multifunctionality of carriers, making it difficult to achieve highly sensitive and stable detection and measurement.
Innovation Solution
A method involving the specific binding of an avidin compound to a biotinylated label, where probes with thiol groups are first bound to avidin and then to a water-soluble carrier, allowing for the formation of a highly sensitive and stable complex with favorable reproducibility, using a strong affinity between avidin and biotin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If probes are polymerized to increase molecular weight and label binding capacity, then sensitivity is improved, but polymerization control becomes difficult
Solution Approach 1:
The patent introduces a carrier molecule as an intermediary component to which multiple probes are attached. This carrier serves as a mediator that enables controlled assembly of probe-label conjugates without requiring direct polymerization of the probes themselves, thus maintaining sensitivity while improving manufacturability
Solution Approach 2:
The probe-label conjugate is segmented into distinct functional components: probes attached to a carrier, and labels attached to the probes. This segmentation allows independent optimization and control of each component, making the overall system easier to manufacture while maintaining high sensitivity
2Measurement precision
If high molecular weight probe-label conjugates are used to increase label capacity, then sensitivity is improved, but reactivity control becomes difficult
Solution Approach 1:
The patent applies local quality by concentrating label molecules at specific locations on the carrier surface where probes are attached. This localized arrangement ensures that high label capacity is achieved only where needed for detection, while maintaining controlled reactivity throughout the conjugate structure
3Power
If carriers with multifunctionality are used to bind probes and labels, then reactivity is improved, but blank value reaction increases
Solution Approach 1:
The patent extracts and separates the binding functions into distinct steps: first binding probes to the carrier, then binding labels to the probes. This separation removes the harmful effect of simultaneous multifunctional binding that causes blank value reactions, while maintaining the reactivity benefits of carrier-based assembly
4Power
If carriers are first bound to probes or labels in complex formation, then reactivity is improved, but reproducibility decreases
Solution Approach 1:
The patent applies preliminary action by pre-attaching probes to the carrier in a controlled first step, creating a stable intermediate structure. This preliminary arrangement ensures consistent geometry and spacing before label attachment, improving reproducibility while maintaining the reactivity advantages of carrier-based conjugates
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
This approach enables the production of a high-sensitive and stable complex of labeled probes and a water-soluble carrier that specifically binds to target substances, enhancing detection and measurement sensitivity and stability while minimizing non-specific reactions.
Implementation Method 1
the specific binding of an avidin compound to a biotinylated label, where probes with thiol groups are first bound to avidin and then to a water-soluble carrier, allowing for the formation of a highly sensitive and stable complex with favorable reproducibility, using a strong affinity between avidin and biotin
Implementation Method 2
binding a probe having one or more thiol groups, being capable of binding to a target substance, to an avidin compound having one or more maleimide groups to obtain a probe conjugate
Data Source
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AI summary
The purpose is to produce, with high reproducibility, a complex of labeled probes and a carrier, said complex being to be used for detecting and measuring a target substance to be measured with high sensitivity and high stability. The means for accomplishing the purpose is that a label is bound to a probe-water soluble carrier conjugate using specific binding of an avidin compound such as avidin, streptavidin, etc. to biotin, and the binding of the avidin compound to the probe is performed before the binding to the carrier. Namely, after conjugating the avidin compound to a substance which is capable of binding to the target substance, the conjugate is bound to a high-molecule water-soluble carrier to produce a complex of the avidinized probes and the water-soluble carrier. Then the complex of the avidinized probes and the water-soluble carrier is mixed with a biotinylated label. Thus, a stable complex of the labeled probes and the water-soluble carrier, which enables the highly sensitive detection and measurement of the target substance, can be obtained with high reproducibility via the specific binding of the avidin compound to biotin.