TY  - JOUR
A1  - Heinsohn, Natascha Katharina
A1  - Niedl, Robert Raimund
A1  - Anielski, Alexander
A1  - Lisdat, Fred
A1  - Beta, Carsten
T1  - Electrophoretic mu PAD for purification and analysis of DNA samples
JF  - Biosensors : open access journal
N2  - In this work, the fabrication and characterization of a simple, inexpensive, and effective microfluidic paper analytic device (mu PAD) for monitoring DNA samples is reported. The glass microfiber-based chip has been fabricated by a new wax-based transfer-printing technique and an electrode printing process. It is capable of moving DNA effectively in a time-dependent fashion. The nucleic acid sample is not damaged by this process and is accumulated in front of the anode, but not directly on the electrode. Thus, further DNA processing is feasible. The system allows the DNA to be purified by separating it from other components in sample mixtures such as proteins. Furthermore, it is demonstrated that DNA can be moved through several layers of the glass fiber material. This proof of concept will provide the basis for the development of rapid test systems, e.g., for the detection of pathogens in water samples.
KW  - microfluidic paper analytic device (mu PAD)
KW  - patterning glass microfiber
KW  - fiber-electrophoresis chip
KW  - DNA
KW  - imprinted electrodes
KW  - cross layer chip
KW  - polymerase chain reaction (PCR)
KW  - purification
Y1  - 2022
U6  - https://doi.org/10.3390/bios12020062
SN  - 2079-6374
VL  - 12
IS  - 2
PB  - MDPI
CY  - Basel
ER  - 
TY  - JOUR
A1  - Sagu Tchewonpi, Sorel
A1  - Huschek, Gerd
A1  - Waldbach Braga, Tess
A1  - Rackiewicz, Michal
A1  - Homann, Thomas
A1  - Rawel, Harshadrai Manilal
T1  - Design of Experiment (DoE) for Optimization of HPLC Conditions for the Simultaneous Fractionation of Seven α-Amylase/Trypsin Inhibitors from Wheat (Triticum aestivum L.)
JF  - Processes : open access journal
N2  - Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of the limiting problems. In this work, a simplified approach based on the successive fractionation of ATI extracts by reverse phase and ion exchange chromatography was developed. ATIs were first extracted from wheat flour using a combination of Tris buffer and chloroform/methanol methods. The separation of the extracts on a C18 column generated two main fractions of interest F1 and F2. The response surface methodology with the Doehlert design allowed optimizing the operating parameters of the strong anion exchange chromatography. Finally, the seven major wheat ATIs namely P01083, P17314, P16850, P01085, P16851, P16159, and P83207 were recovered with purity levels (according to the targeted LC-MS/MS analysis) of 98.2 ± 0.7; 98.1 ± 0.8; 97.9 ± 0.5; 95.1 ± 0.8; 98.3 ± 0.4; 96.9 ± 0.5, and 96.2 ± 0.4%, respectively. MALDI-TOF-MS analysis revealed single peaks in each of the pure fractions and the mass analysis yielded deviations of 0.4, 1.9, 0.1, 0.2, 0.2, 0.9, and 0.1% between the theoretical and the determined masses of P01083, P17314, P16850, P01085, P16851, P16159, and P83207, respectively. Overall, the study allowed establishing an efficient purification process of the most important wheat ATIs. This paves the way for further in-depth investigation of the ATIs to gain more knowledge related to their involvement in NCWS disease and to allow the absolute quantification in wheat samples.
KW  - wheat
KW  - α-amylase/trypsin inhibitors
KW  - fractionation
KW  - purification
KW  - reversed-phase chromatography
KW  - ion-exchange chromatography
KW  - design of experiment
KW  - LC–MS/MS
KW  - MALDI-TOF-MS
Y1  - 2022
U6  - https://doi.org/10.3390/pr10020259
SN  - 2227-9717
VL  - 10
SP  - 1
EP  - 18
PB  - MDPI
CY  - Basel, Schweiz
ET  - 2
ER  -