Miniaturized Hemagglutinin Complex Protein for E-cadherin Inhibition
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Solution Overview
Problem
Current methods for maintaining undifferentiated pluripotent stem cells are complex and labor-intensive, requiring manual removal of deviated cells under a microscope, and existing miniaturized hemagglutinin (HA) complexes lack cadherin function inhibitory activity.
Innovation Solution
A novel miniaturized HA complex protein is developed, which maintains cadherin function inhibitory activity by deleting parts of the HA1 and HA3 subcomponents, utilizing the HA2-HA3 connecting region for binding with E-cadherin, and using a linker to fuse HA2 and HA3 subcomponents, thereby simplifying production and enhancing handling.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the whole HA complex is produced recombinantly, then cadherin function inhibitory activity is maintained, but production complexity and time increase due to multiple subunit expression and purification steps
Solution Approach 1:
The invention divides the HA complex into separate subunit proteins (HA1, HA2, HA3) that can be independently expressed and purified, then reconstituted into the functional complex. This segmentation allows each subunit to be produced separately using different expression systems optimized for their specific properties, reducing overall production complexity while maintaining the integrity and activity of the final complex.
Solution Approach 2:
The invention combines multiple separately produced subunit proteins (HA1 dimer, HA2, and HA3) in a controlled assembly process to form the functional HA complex. By merging these individually optimized components, the invention achieves both production efficiency (each subunit can be produced independently) and functional reliability (the assembled complex maintains cadherin inhibitory activity).
2Ease of manufacture
If miniaturized HA complex is used, then production is simplified and handling is easier, but cadherin function inhibitory activity is lost
Solution Approach 1:
The invention extracts and retains only the essential functional regions of the HA complex subunits that are necessary for cadherin binding and inhibitory activity. By identifying and preserving these critical domains while removing non-essential portions, the invention creates a miniaturized version that maintains biological function while achieving production and handling advantages.
Solution Approach 2:
The invention applies local quality by maintaining the full sequence and structure of specific critical regions (such as the HA2-HA3 connecting region that binds E-cadherin) while miniaturizing other portions of the complex. This selective preservation of local functional quality ensures that the miniaturized complex retains cadherin inhibitory activity despite overall size reduction.
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 miniaturized HA complex protein effectively inhibits E-cadherin function, facilitating the maintenance of undifferentiated pluripotent stem cells by removing deviated cells efficiently, while also promoting cell proliferation and simplifying the production process.
Implementation Method 1
the HA2-HA3 connecting region for binding with E-cadherin
Data Source
AI summary
A nucleic acid encoding a miniatureized hemagglutinin complex protein, wherein the miniaturized hemagglutinin complex protein has function inhibitory activity against E-cadherin, and wherein the miniaturized hemagglutinin complex protein is selected from the group consisting of: (1) mini-HA consisting of HA1 subcomponent of SEQ ID NO: 10, HA2 subcomponent of SEQ ID NO: 4, and HA3 subcomponent fragment of SEQ ID NO: 2, (2) FL(2+3)-C consisting of HA2 subcomponent of SEQ ID NO: 4 and HA3 component with Strep-Tag (C-terminus) of SEQ ID NO: 44, (3) linked-mini (2+3)/BB of SEQ ID NO: 47, (4) linked-mini (2+3)/CB of SEQ ID NO: 48, (5) linked-mini (2+3)/CB-YFDY of SEQ ID NO: 49, and (6) linked-mini (2+3)/CB-LD/YFDY of SEQ ID NO: 50.