Monoclonal Colony Selection via In-Situ Binding Complex Detection
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Solution Overview
Problem
Current methods for selecting monoclonal cell colonies that secrete a product of interest are cumbersome due to the need to take and analyze aliquots from each well, complicating the assessment of secretion and colony formation in multi-well plate systems.
Innovation Solution
A method involving immobilized first binding agents on assessment surfaces within cell culture spaces, combined with non-immobilized second binding agents that bind to the product of interest, allows for the localization and detection of secreted products without removing aliquots, enabling the selection of monoclonal cell colonies that secrete specific proteins or antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If aliquots are taken from each well for analysis, then secretion of product of interest can be assessed, but the method becomes complicated and time-consuming
Solution Approach 1:
The patent introduces binding agents as intermediaries that capture the product of interest at the well bottom surface. The first binding agent (immobilized) and second binding agent (in solution) form a detectable complex that serves as a mediator between the secreted product and the detection system, eliminating the need for aliquot removal while enabling precise secretion assessment
Solution Approach 2:
The patent replaces the mechanical operation of taking and analyzing aliquots with an in-situ binding and detection system. Instead of physically removing and analyzing samples, the system uses binding agents to capture and localize the product of interest at the well bottom, substituting mechanical sample handling with a chemical binding-based detection approach
2Measurement precision
If aliquots are taken from each well for analysis, then product of interest can be detected, but productivity is reduced due to additional steps
Solution Approach 1:
The binding agents act as intermediaries that enable direct detection of the product of interest in the well without requiring sample removal. The first binding agent immobilized on the well bottom and the second binding agent in solution form a detectable complex that provides precise detection while maintaining productivity by eliminating time-consuming aliquot handling steps
Solution Approach 2:
The system enables self-service detection where the binding agents automatically capture and localize the product of interest at the well bottom surface. The secreted product naturally binds to the immobilized first binding agent, and the added second binding agent forms a detectable complex, eliminating the need for external sample processing operations
3Measurement precision
If white light imaging is used to assess colony formation, then monoclonal colonies can be identified, but secretion assessment still requires separate aliquot analysis
Solution Approach 1:
The patent merges colony formation assessment and secretion assessment into a single integrated process. Both assessments are performed by imaging the well bottom surface, where the binding complex localizes the secreted product at the same location where colonies grow, allowing simultaneous evaluation of both parameters without separate aliquot analysis
Solution Approach 2:
The well bottom surface serves multiple functions: it supports cell colony growth, provides a platform for binding agent immobilization, and acts as a detection surface for both colony morphology and secreted product localization. This multi-functional design enables unified assessment of both colony formation and secretion through a single imaging approach
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 simplifies the identification and selection of monoclonal cell colonies by localizing and detecting secreted products at the assessment surface, reducing complexity and improving efficiency in identifying high-producing colonies without disrupting the culture medium or cells.
Implementation Method 1
providing each said assessment surface with a first binding agent that is immobilised on the assessment surface; wherein each of the first and second binding agents binds to the product of interest
Implementation Method 2
providing a second binding agent in the cell culture spaces, the second binding agent not being immobilised; wherein each of the first and second binding agents binds to the product of interest so that secretion of the product of interest by cells in any one of the cell culture spaces leads to a complex comprising the first binding agent, the product of interest and the second binding agent forming at the corresponding assessment surface
Data Source
AI summary
A method of selecting a monoclonal cell colony that secretes a product of interest providing a plurality of cell culture spaces; each cell culture space has a respective assessment surface with a first binding agent immobilized on the assessment surface. The cells are incubated in a growth medium in the cell culture spaces for replication of the cells. At least some of the cells secrete a product of interest. A second binding agent is provided in the cell culture spaces. Initially, the second binding agent is not immobilized. Each of the first and second binding agents binds to the product of interest so that secretion of the product of interest by cells in any one of the cell culture spaces leads to a complex with the first binding agent, the product of interest and the second binding agent forming at the corresponding assessment surface of the cell culture space.