Lecture
Nucleic Acid Extraction Through Zwitterionic Nanofibers Integrated in Flow-Through Micro-Paper-Based Devices
- 10.04.2024 at 13:30 - 14:00
- ICM Saal 4b
- Language: English
- Type: Lecture
Lecture description
Sensitive and yet rapid detection of pathogens such as HIV, Malaria and SARS CoV-2 rely on the identification and quantification of their nucleic acids (NAs), enabling the detection early on in the infection cycle. The extraction of nucleic acids from the biological matrices is mandatory to allow for subsequent sensitive and specific molecular detection or amplification strategies [1]. Various extraction technologies, e.g. spin-columns and FTA cards, are commercially available and offer high efficiency but are unsuitable for point-of-care (POC) applications due to the need for equipment, high costs. Therefore, the development of new NAs extraction methods that allow for in-line isolation for a sample-to-answer system are necessary [2].
Owing to their high surface-to-volume ratio, light-weight, tunable chemical properties and ease of mass production, nanofibers (NFs) are a promising candidate to be integrated as an in-line NA extractor in various paper-based analytical devices (PADs) for POC applications. Here, we developed novel electrospun zwitterionic NFs made from nylon doped with the cationic charged poly(allylamine hydrochloride) (PAH) and polyanionic polymer poly(acrylic acid) (PAA). Simply by changing the pH, these NFs can either efficiently adsorb or elute NAs. The extraction efficiency is strongly influenced by the composition of the polymers as well as fiber morphology, where a correlation between the number of nanonets within the fiber mat and the extraction efficiency was revealed. Under optimal conditions, over 90% adsorption and up to 80% elution efficiency within a rapid timeframe of under 30 minutes was achieved without the need for additional devices. A long-term stability study demonstrated that the full functionality of zwitterionic NFs could be preserved over months.
The as-developed zwitterionic NFs were further integrated into PADs enabling an efficient flow-through extraction of NAs. Due to their high surface area with immense porosity, the NFs in the flow system exhibit a comparable extraction efficiency even in the absence of any incubation time, allowing a direct coupling of the extraction with an on-chip amplification strategy, such as RPA, followed by a detection. Furthermore, initial interference studies indicated minimal serum protein adsorption (below 40%) on the NFs, with only about 10% eluted along with NAs. With these combined advantages of a cheap and lightweight material, as well as the rapid, efficient and in-line extraction feasibility those zwitterionic NFs hold a great promise for future application in the POC detection of pathogens in resource limited settings.
Literature:
[1] Niemz, A, Tends Biotechnol 2011, 29, 5. [2] Ali, N, Biomed Res. Int. 2017, 2017
Owing to their high surface-to-volume ratio, light-weight, tunable chemical properties and ease of mass production, nanofibers (NFs) are a promising candidate to be integrated as an in-line NA extractor in various paper-based analytical devices (PADs) for POC applications. Here, we developed novel electrospun zwitterionic NFs made from nylon doped with the cationic charged poly(allylamine hydrochloride) (PAH) and polyanionic polymer poly(acrylic acid) (PAA). Simply by changing the pH, these NFs can either efficiently adsorb or elute NAs. The extraction efficiency is strongly influenced by the composition of the polymers as well as fiber morphology, where a correlation between the number of nanonets within the fiber mat and the extraction efficiency was revealed. Under optimal conditions, over 90% adsorption and up to 80% elution efficiency within a rapid timeframe of under 30 minutes was achieved without the need for additional devices. A long-term stability study demonstrated that the full functionality of zwitterionic NFs could be preserved over months.
The as-developed zwitterionic NFs were further integrated into PADs enabling an efficient flow-through extraction of NAs. Due to their high surface area with immense porosity, the NFs in the flow system exhibit a comparable extraction efficiency even in the absence of any incubation time, allowing a direct coupling of the extraction with an on-chip amplification strategy, such as RPA, followed by a detection. Furthermore, initial interference studies indicated minimal serum protein adsorption (below 40%) on the NFs, with only about 10% eluted along with NAs. With these combined advantages of a cheap and lightweight material, as well as the rapid, efficient and in-line extraction feasibility those zwitterionic NFs hold a great promise for future application in the POC detection of pathogens in resource limited settings.
Literature:
[1] Niemz, A, Tends Biotechnol 2011, 29, 5. [2] Ali, N, Biomed Res. Int. 2017, 2017
All lectures within this session
- 12:30 - 13:00Standard Addition for Immunoassays - New Possibilities for Quantitative Lateral Flow Tests
- 13:00 - 13:30From Complex to Simple: New Compact Lateral Flow Test Systems
- 13:30 - 14:00Nucleic Acid Extraction Through Zwitterionic Nanofibers Integrated in Flow-Through Micro-Paper-Based Devices
- 14:00 - 14:30Next Generation of Unisart® Membranes – Multiplexing with Unisart StructSure®