Discover how patent US 2017/0260530 A1 introduces synthetic DNA aptamers that transform gelsolin detection and purification, eliminating the need for animal-derived reagents while delivering superior performance and cost-effectiveness.
Patent Application Overview
Purpose & Significance
Quantify Gelsolin
Novel DNA aptamers enable precise measurement of gelsolin levels in biological samples, offering enhanced sensitivity and reproducibility.
Purify Protein
Aptamers create efficient affinity matrices for purifying gelsolin and its variants, including difficult tagless forms.
Animal-Free Alternative
Replaces traditional animal-derived reagents with synthetic DNA aptamers, addressing ethical concerns and reducing costs.
Background & Challenges
Current Methods
Rely on actin extracted from rabbit/chicken muscle or anti-gelsolin antibodies produced in animals.
Expensive and labor-intensive
Variable batch-to-batch quality
Ethical concerns regarding animal use
Gelsolin Function
Six-domain protein (G1-G6) with three human isoforms that performs crucial roles:
Actin filament depolymerization
Nucleation of new filaments
Clearing toxic actin from plasma
Key Patent Claims
Novel DNA Aptamers
Unique nucleotide sequences (SEQ ID Nos. 6-9) with specific binding properties to gelsolin.
Binding Specificity
Aptamers bind to full-length gelsolin and its N-terminal (G1-G3) and C-terminal (G4-G6) variants.
Quantification Methods
Two approaches using biotin-labeled aptamers with streptavidin and HRP for colorimetric detection.
Purification Applications
Claims utility in isolating gelsolin from complex biological samples with high specificity.
Technical Specifications
Detection Methods
Colorimetric readout at 450nm after HRP substrate reaction
Aptamer Sequences
Four distinct sequences (75-77 bases) with specific binding domains
Production Methods
Synthesized via PCR, chemical synthesis, or bacterial production
Application Platforms
Adaptable to various detection and purification systems
Scientific & Practical Implications
Diagnostics
Enables quantification of plasma gelsolin as potential biomarker for conditions involving cell necrosis (trauma, sepsis).
Scalability
Mass production via PCR or bacterial systems offers significant cost advantages over antibody production.
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Research
Provides reliable tools for studying gelsolin's role in cellular processes and disease mechanisms.
Therapeutics
Facilitates production of purified gelsolin for potential therapeutic applications.
Limitations & Future Directions
Experimental Validation Gaps
While claims suggest specificity and sensitivity comparable to traditional methods, detailed experimental results regarding binding affinities and detection limits are not fully presented.
Missing Visual Data
References to Figures 1-6 appear in the text but aren't included in the excerpts, preventing assessment of binding curves, purification yields, and other critical visual data.
Comparative Performance
More comprehensive benchmarking against established methods would strengthen the case for widespread adoption of these novel aptamers in research and diagnostic applications.