Modified LAMP-2B Plasmid for Stable Biomolecule Delivery
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Solution Overview
Problem
Current methods for the stable expression and delivery of biomolecules face challenges in achieving high stability and targetability, particularly when using conventional lysosome-associated membrane glycoprotein 2B (LAMP-2B), as they are prone to lysosomal degradation and have limited delivery efficiency to target cells.
Innovation Solution
A plasmid platform is developed that removes the intracellular and extracellular domains of LAMP-2B, incorporating a nucleic acid sequence encoding a modified protein and a glycosylated region to enhance stability and delivery efficiency, utilizing exosomes for targeted biomolecule delivery, including cancer diagnosis and treatment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional LAMP-2B is used for biomolecule delivery, then the delivery system can be constructed, but the biomolecules are prone to lysosomal degradation and have limited delivery efficiency
Solution Approach 1:
The patent removes the intracellular domain (IC) and/or extracellular domain (EC) of LAMP-2B to eliminate the problematic lysosomal targeting function while preserving the membrane anchor function. This extraction of specific domains resolves the contradiction by eliminating the degradation pathway (improving reliability) while maintaining delivery capability (preserving productivity).
Solution Approach 2:
The patent modifies specific regions of LAMP-2B by selectively removing only the IC and/or EC domains while retaining the transmembrane domain and cytoplasmic tail. This localized modification approach allows the protein to maintain essential functions (membrane anchoring, exosome association) while eliminating harmful lysosomal targeting, thus resolving the contradiction between stability and delivery efficiency.
2Stability of the object's composition
If the intracellular domain of LAMP-2B is removed to prevent lysosomal degradation, then biomolecule stability increases, but protein structure完整性 may be compromised
Solution Approach 1:
The patent extracts only the intracellular domain (amino acids 26-106) from LAMP-2B while retaining the transmembrane domain (amino acids 107-136) and cytoplasmic tail (amino acids 137-217). This selective extraction maintains the essential structural components needed for membrane anchoring and exosome association while eliminating the lysosomal degradation pathway, thus resolving the contradiction between stability and structural integrity.
Solution Approach 2:
The modification is applied locally to specific domains of LAMP-2B rather than the entire protein structure. By removing only the IC and/or EC domains while preserving the transmembrane and cytoplasmic regions, the patent maintains local structural integrity where needed while achieving global stability improvement.
3Productivity
If exosomes are used for targeted delivery, then delivery efficiency to cancer cells increases, but the system complexity increases
Solution Approach 1:
The patent utilizes the cell's natural exosome secretion machinery to deliver the modified LAMP-2B protein and associated biomolecules. The exosomes self-assemble and self-deliver to target cells using endogenous pathways, eliminating the need for complex external delivery systems or artificial targeting mechanisms, thus resolving the contradiction between delivery efficiency and system complexity.
Solution Approach 2:
The modified LAMP-2B protein serves multiple functions: it acts as a membrane anchor, associates with exosomes, and enables targeted delivery to cancer cells. This multi-functionality reduces the need for separate components or mechanisms, simplifying the overall system while maintaining high delivery efficiency.
Data Source
AI summary
A plasmid platform according to an embodiment includes a nucleic acid sequence encoding a modified protein from which an intracellular domain, an extracellular domain, or a combination thereof of lysosome-associated membrane glycoprotein 2B (LAMP-2B) has been removed, and is based on the discovery that removing a specific domain of LAMP-2B significantly stabilizes the expression of intended biomolecules and significantly increases the efficiency with which intended biomolecules are delivered. The plasmid platform is expected to provide high technical utility in the expression and delivery of proteins of interest, particularly expression and delivery using exosomes, and may be expanded to various associated technical fields or uses.