Feeder Cell Line for Direct Monoclonal Antibody Isolation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating monoclonal antibodies using Epstein-Barr virus (EBV) are inefficient, require multiple subcloning steps, and are costly and time-consuming, while single-cell molecular cloning methods are expensive and lack pre-expression antibody testing capabilities.
Innovation Solution
A novel feeder cell line, Amalthea, expressing mega CD40 ligand, CD23, and a fluorescent protein, supports monoclonal antibody-producing B cells, enabling direct culture and isolation without subcloning, and allows for immediate antibody assessment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If EBV is used to isolate monoclonal antibodies, then antibody production is achieved, but transformation efficiency is low and multiple subcloning steps are required
Solution Approach 1:
The patent introduces a feeder cell line as an intermediary component that provides essential growth factors and support signals to EBV-transformed B cells. This feeder layer enables direct monoclonal outgrowth without requiring multiple subcloning steps, resolving the contradiction between transformation efficiency and procedural complexity
Solution Approach 2:
The feeder cell line is pre-engineered to express specific surface molecules and growth factors that create an optimal microenvironment for B cell transformation and monoclonal outgrowth. This preliminary preparation eliminates the need for subsequent subcloning operations, improving both efficiency and reducing complexity
2Productivity
If single-cell molecular cloning is used, then throughput is increased and time is reduced, but cost increases significantly
Solution Approach 1:
The patent uses a feeder cell line system that copies and amplifies the successful B cell clone directly in culture, eliminating the need for expensive molecular cloning procedures. This biological copying approach achieves high throughput while maintaining cost-effectiveness
Solution Approach 2:
The feeder cell line serves as a disposable support system that enables direct monoclonal outgrowth without requiring expensive molecular cloning infrastructure. This approach replaces costly single-cell sorting and molecular biology facilities with a simpler, more economical cell culture system
3Reliability
If classic EBV method is used, then antibody production is achieved, but time required is several weeks
Solution Approach 1:
The feeder cell line is pre-conditioned with growth factors and surface molecules that immediately support B cell transformation and monoclonal outgrowth. This preliminary preparation accelerates the entire process from several weeks to days, while maintaining reliable antibody production
Solution Approach 2:
The feeder cell line provides continuous support signals and growth factors throughout the culture period, enabling uninterrupted monoclonal outgrowth and antibody production. This continuous action eliminates the need for sequential subcloning steps, significantly reducing the time required while maintaining production reliability
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 Amalthea feeder cell line significantly increases transformation efficiency, reduces costs by 50-100-fold, and enhances throughput, allowing for rapid antibody production and testing.
Implementation Method 1
EBV induces the continuous proliferation (transformation and 'immortalisation') of human B cells to create lymphoblastoid cell lines (LCLs)
Implementation Method 2
the feeder cell line expresses: a fluorescent protein that is not expressed by the B cell
Data Source
AI summary
The invention relates to monoclonal antibody production and isolation, and particularly, although not exclusively, to a novel feeder cell line for culturing a monoclonal antibody-producing B cell. The invention also extends to the use of the feeder cell line in culturing a monoclonal antibody-producing B cell, and isolating the B cell from the cell culture media. The invention further extends to methods for culturing and isolating a monoclonal antibody-producing B cell, as well as a method for isolating a monoclonal antibody.