Recombinant Lectin Solubility via Amino Acid Modification
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Solution Overview
Problem
Native Sclerotium rolfsii lectin has solubility problems that hinder its practical applications in medical assays and therapies, limiting its effectiveness as a diagnostic and therapeutic agent for cancer.
Innovation Solution
Development of recombinant lectins (UC-SS/CSR-1803 and UC-SS/CSR-1.805) with modified amino acid sequences, expressed in E.coli, which exhibit improved solubility and stability, specifically binding to O-glycosidically linked glycans on cancer cells and showing strong affinity for desialofetuin, allowing for enhanced antiproliferative activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If native Sclerotium rolfsii lectin is used, then it shows high binding affinity to cancer cells and antiproliferative activity, but it has solubility problems that hinder practical applications
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the native lectin protein to create recombinant variants (UC-SS/CSR-1803 and UC-SS/CSR-1.805) with altered physicochemical properties. These modifications improve solubility and stability while preserving the carbohydrate-binding activity and antiproliferative effects against cancer cells.
2Ease of manufacture
If recombinant lectins with modified amino acid sequences are developed, then solubility and stability are improved, but production complexity increases
Solution Approach 1:
The patent uses copying by creating recombinant DNA copies of the modified lectin gene sequences and expressing them in E. coli host cells. This allows production of multiple copies of the improved lectin protein through bacterial fermentation, simplifying large-scale manufacturing while maintaining the desired solubility and stability characteristics.
3Productivity
If lectin genes are synthesized and cloned in E.coli, then ease of production is improved, but manufacturing precision requirements increase
Solution Approach 1:
The patent replaces traditional protein purification and characterization methods with molecular biology techniques. By synthesizing lectin genes and cloning them into E. coli, the process substitutes chemical and physical purification steps with genetic engineering approaches, improving productivity while requiring precise control of DNA sequencing and cloning procedures.
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 recombinant lectins demonstrate improved solubility and stability, maintaining strong binding affinity to cancer cells, thereby enhancing their diagnostic and therapeutic potential as antitumor agents with increased ease of production and application.
Implementation Method 1
Lectins can bind reversibly with monosaccharides or the sugar moiety of, glycoproteins or glycolipids or polysaccharides. Some of the lectins have ability to specifically recognize and bind to these altered structures and exerts various physiological effects
Implementation Method 2
The recombinant lectins expressed as soluble proteins and are purified by ion exchange and gel filtration chromatographic techniques
Implementation Method 3
The recombinant lectins expressed as soluble proteins and are purified by ion exchange and gel filtration chromatographic techniques
Data Source
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AI summary
The present invention discloses genes for new variants in E.coli that are more close to the wild type lectin sequence but still differ in surface charges. This recombinant version (exemplified Rec-2 and Rec-3) are cancer cell binding with anti tumor activity show blood group specificity similar to the native lectin but retains the stability and the solubility properties.