DNA-Barcoding Reagent for Antibody Labeling
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for DNA-barcoding antibodies are time-consuming and inefficient, requiring long reaction times and multiple protein concentration and purification steps, making them costly and inaccessible to many biological laboratories.
Innovation Solution
A novel DNA-barcoding reagent with an affinity moiety specific to antibodies, covalently linked to one or more DNA sequences via a linker, allowing for rapid and quantitative attachment to antibodies, enabling complete labeling in under 10 minutes at room temperature, and subsequent scavenging of excess reagent using non-specific immunoglobulins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If existing DNA-barcoding methods are used, then antibodies can be labeled with DNA sequences, but the process requires long reaction times (>16 hours) and multiple purification steps
Solution Approach 1:
The invention changes the chemical parameters of the conjugation reaction by using heterobifunctional crosslinkers with specific reactivity patterns (e.g., NHS ester for amines, maleimide for thiols) and optimizing pH, temperature, and molar ratios to achieve complete labeling in under 10 minutes at room temperature
Solution Approach 2:
The invention performs preliminary activation of the crosslinker reagent and prepares the DNA-barcoding reagent in advance with pre-attached linkers and DNA sequences, so that the actual conjugation step only requires mixing and brief incubation without time-consuming in-situ activation
2Manufacturing precision
If existing DNA-barcoding methods are used, then antibodies can be labeled with DNA sequences, but multiple protein concentration and purification steps are required
Solution Approach 1:
The invention extracts and removes the time-consuming purification steps from the conventional workflow by using crosslinkers with irreversible bonding that prevents unreacted DNA from detaching, and by using scavenging proteins to capture excess reagent, eliminating the need for multiple HPLC or affinity purification steps
Solution Approach 2:
The invention uses disposable scavenging proteins (non-specific immunoglobulins) that can be added in excess to capture unreacted DNA-barcoding reagent, then removed in a single simple filtration or dilution step, replacing expensive and complex multi-step purification protocols
3Manufacturing precision
If existing DNA-barcoding methods are used, then antibodies can be labeled with DNA sequences, but the process is costly and inaccessible to many biological laboratories
Solution Approach 1:
The invention makes the DNA-barcoding process self-service by providing a reagent system that works under physiological conditions (room temperature, aqueous buffer) with simple mixing and brief incubation, requiring no specialized equipment or expertise in complex bioconjugation chemistry, thus making it accessible to any biological laboratory
Solution Approach 2:
The invention creates a universal DNA-barcoding reagent system that can be applied to any antibody with appropriate amino groups or thiol groups, using the same heterobifunctional crosslinker chemistry and scavenging approach, eliminating the need for method development for each specific antibody and making the process broadly accessible
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 method enables cost-efficient and user-friendly DNA-barcoding of antibodies, facilitating multiplexed molecular imaging and protein quantification, and can be used directly in various biotechnological applications without further purification, improving the efficiency and accessibility of DNA-barcoding processes.
Implementation Method 1
an affinity moiety specific towards antibodies, and covalently linked to one or more DNA sequences
Data Source
AI summary
Provided are DNA-barcoding reagents, DNA-barcoded antibodies, and methods of using DNA-barcoding reagents, and DNA-barcoded antibodies. The DNA-barcoding reagents comprise an affinity moiety that is specific towards antibodies, and covalently linked to one or more DNA sequences, optionally, by a linker. Also provided are methods of using the DNA-barcoded antibodies, for example, multiplexed tissue antigen imaging and profiling, multiplexed biomolecule detection, and affinity purification and sorting of marker-positive cells.