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Immobilization of DNA on microporous PVDF membranes by plasma polymerization
Dar-Jong Lin1, Dong-Tsamn Lin, Tai-Horng Young
1Department of Chemical and Materials Engineering, Tamkang University, Taipei 25137, Taiwan.
Abstract:
Microporous poly(vinylidene fluoride) (PVDF) membranes with different porous surface morphologies were prepared by immersion-precipitation from coagulation baths of different strengths. On these membranes single-strand deoxyribonucleic acid (ss-DNA) was covalently immobilized by a dual-step procedure. First, poly(glycidyl methacrylate) (PGMA) was grafted on PVDF membranes by plasma-induced free radical polymerization. Then, ss-DNA was bonded to PGMA through ring-opening reactions of epoxies with amine to form amino alcohols. The highest attainable graft yield of PGMA on PVDF membrane was 0.3 mg/cm(2), which was the case when a highly porous PVDF membrane was employed as the substrate. For immobilization of ss-DNA, the yield was found to depend significantly on the reaction temperature and pH. The maximal value was 48.5 mug/cm(2). Furthermore, adsorption tests of anti-DNA antibody were carried out on membranes with and without immobilized ss-DNA using serum obtained from systemic lupus erythematosus patients. The results indicated that the immobilized DNA could effectively adsorb the antibody in the serum.
Insights
Researchers developed functionalized poly(vinylidene fluoride) (PVDF) membranes for enhanced single-strand deoxyribonucleic acid (ss-DNA) immobilization. These modified membranes effectively captured anti-DNA antibodies from serum, showing promise for diagnostic applications.
Area of Science:
- Materials Science
- Biotechnology
- Chemical Engineering
Background:
- Microporous membranes are crucial for various applications, including biosensing.
- Immobilizing biomolecules like single-strand deoxyribonucleic acid (ss-DNA) onto synthetic membranes is challenging.
- Poly(vinylidene fluoride) (PVDF) membranes offer a potential platform for biomolecule immobilization due to their chemical stability and tunable porosity.
Purpose of the Study:
- To develop a robust method for covalently immobilizing ss-DNA onto PVDF membranes.
- To optimize the grafting and immobilization procedures for high yields.
- To evaluate the performance of ss-DNA immobilized PVDF membranes in capturing specific antibodies.
Main Methods:
- Preparation of microporous PVDF membranes with varying morphologies via immersion-precipitation.
- Plasma-induced grafting of poly(glycidyl methacrylate) (PGMA) onto PVDF membranes.
- Covalent immobilization of ss-DNA onto PGMA-grafted PVDF membranes through ring-opening reactions.
- Characterization of graft yields and optimization of reaction conditions (temperature, pH).
- Adsorption studies using anti-DNA antibodies from systemic lupus erythematosus patient serum.
Main Results:
- Achieved a maximum PGMA graft yield of 0.3 mg/cm(2) on highly porous PVDF membranes.
- Maximal ss-DNA immobilization yield reached 48.5 μg/cm(2), dependent on reaction temperature and pH.
- ss-DNA immobilized membranes demonstrated effective adsorption of anti-DNA antibodies from patient serum.
- Control membranes without immobilized ss-DNA showed negligible antibody adsorption.
Conclusions:
- A successful dual-step procedure for covalent ss-DNA immobilization on PVDF membranes was established.
- The method allows for high biomolecule loading and is sensitive to reaction parameters.
- The functionalized membranes show significant potential for developing sensitive diagnostic tools for autoimmune diseases like systemic lupus erythematosus.
