Surface Protein Ligation Peptide for Biomolecule Cell Selection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for selecting antibody-producing hybridoma cells are inefficient due to reliance on affinity between secreted antibodies and membrane-bound antibody binding proteins, leading to sub-optimal identification and potential damage to cells, and are limited to antibodies, not applicable to other biomolecules.
Innovation Solution
A method involving a surface protein with an extracellular ligation peptide sequence for enzymatic conjugation of an adapter ligand, enabling indirect or direct coupling of a molecular catcher structure with specific binding sites for released biomolecules, allowing for specific binding and detection of biomolecules at the cell surface, independent of their Fc part.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If antibody binding proteins with high affinity to secreted antibodies are used for cell surface marking, then cell identification becomes easier, but optimal antibody-producing cells may be missed due to competitive displacement by sub-optimal antibodies with greater affinity to the binding protein
Solution Approach 1:
The patent introduces an adapter ligand as an intermediary component that couples the molecular catcher structure to the cell surface protein. This adapter system ( comprising ligation peptide sequence, adapter ligand, and molecular catcher structure) mediates between the cell surface and the detection system, allowing specific binding of secreted biomolecules without relying on high-affinity direct binding between antibody binding proteins and antibodies, thereby eliminating competitive displacement issues while maintaining selection accuracy
Solution Approach 2:
The patent segments the cell surface marking system into distinct functional components: a ligation peptide sequence for enzymatic conjugation, an adapter ligand for coupling, and a molecular catcher structure with specific binding sites. This segmentation allows each component to perform its specific function independently, enabling precise selection based on biomolecule affinity without interference from non-specific high-affinity binding
2Ease of manufacture
If chemical biotinylation of cell surfaces is performed to enable capture component binding, then cell surface marking is achieved, but cell vitality is reduced and expression of desired cell products decreases
Solution Approach 1:
The patent replaces the chemical biotinylation method with an enzymatic conjugation system using a ligation peptide sequence that can be enzymatically modified. This substitution eliminates the need for harsh chemical reagents and complex separation steps, thereby preserving cell vitality and maintaining high expression levels of desired cell products while achieving effective cell surface marking
Solution Approach 2:
The ligation peptide sequence is designed to be extracellularly accessible and enzymatically modifiable, allowing the cell surface protein to serve its own marking function without requiring external chemical treatment or separation of cells from growth medium. The system enables self-marking through enzymatic conjugation, avoiding cell stress and damage associated with chemical biotinylation
3Reliability
If antibody binding proteins are used for cell surface marking, then the method is specific to antibodies, but this limits applicability to other types of biomolecules
Solution Approach 1:
The patent creates a universal cell surface marking system where the molecular catcher structure can be configured with specific binding sites for different types of biomolecules. The adapter ligand system serves multiple functions: it couples various catcher structures to the cell surface and enables detection of diverse biomolecules including antibodies, proteins, hormones, and other molecules, making the method universally applicable across different biomolecule types while maintaining 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
Enables reliable and specific selection of cells producing biomolecules with optimal affinity to target molecules, ensuring cell vitality and broad applicability beyond antibodies, including proteins, hormones, and other biomolecules.
Implementation Method 1
a surface protein having an extracellularly exposed ligation peptide sequence for enzymatic conjugation of an adapter ligand
Implementation Method 2
said adapter ligand being suitable for the indirect or direct coupling of a molecular catcher structure which has at least one specific binding site for the released biomolecules
Implementation Method 3
specific binding of at least one of the released biomolecules to the specific binding site
Data Source
AI summary
The invention relates, firstly, to a biomolecule-releasing cell characterised by a surface protein with an extracellularly exposed ligation peptide sequence for enzymatic conjugation of an adapter ligand, said adapter ligand being suitable for the indirect or direct coupling of a molecular catcher structure which has at least one specific binding site for the released biomolecules and which is at a distance from the specific binding site, secondly, to a method for selecting such a cell, and thirdly, to a means for this method comprising the surface protein that is characterised by an extracellularly exposable ligation peptide sequence for enzymatic conjugation of an adapter ligand, a nucleic acid coding for the surface protein, an expression vector for the nucleic acid, and a cell containing the expression vector.