Removal of urea from urea-rich protein samples using metal ions in a microfluidic device |
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| Affiliation: | 1. Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea;2. Department of Chemical and Biological Engineering Chungju National University, 123 Geomdan-ri, Iryu-myeon, Chungju, Chungbuk 380-702, Republic of Korea;1. Department of Microbiology, College of Medicine, Hallym University, 1 Okcheon-dong, Chuncheon, Gangwon-do 200-702, Republic of Korea;2. Ilsong Institute of Life Science, Hallym University, 1605-4 Gwanyang-dong, Dongan-gu, Anyang, Gyeonggi-do 431-060, Republic of Korea;3. Department of Food Science and Nutrition, Pukyong National University, 599-1 Daeyeon-3-dong, Nam-gu, Busan 608-737, Republic of Korea;4. New York State Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, NY 10314, USA;3. Department of Microbiology, University of Illinois, Urbana, Illinois 61801;6. Institute for Genomic Biology, University of Illinois, Urbana, Illinois 61801;4. Department of Biology and Biochemistry, University of Houston, Houston, Texas 77204;5. Department of Microbiology, Immunology, and Molecular Genetics, UCLA, Los Angeles, California 90095;1. State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;2. Shanghai Institute of Medical Genetics, Shanghai Children''s Hospital, Shanghai Jiao Tong University, Shanghai 200040, China;3. Key Laboratory of Embryo Molecular Biology, Ministry of Health, and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai, PR China;1. Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061, USA;2. Department of Chemistry, Faculty of Science, Damietta University, Damietta, 34517, Egypt;3. Department of Food Technology, Arid Lands Cultivation Research Institute, City of Scientific Research and Technological Applications, Alexandria, 21934, Egypt;4. Fralin Life Science Institute, Virginia Tech, Blacksburg, VA 24061, USA;5. Virginia Tech Center for Drug Discovery, Virginia Tech, Blacksburg, VA 24061, USA;1. Graduate Program in Cell, Molecular, and Structural Biology, Miami University, Oxford, OH, 45056 USA;2. Department of Microbiology, Miami University, Oxford, OH, 45056 USA |
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| Abstract: | Urea is commonly used to lyse cultured cells and solubilize proteins from a biological source. In this study, after extracting biomolecules using a lysis buffer that included urea for an effective cleaning of protein from a urea-rich protein sample, a five-flow microfluidic desalting system was applied using the metal ions of Mn2+, Zn2+ and Fe3+, which have urea affinity-capturing properties. This device effectively removed urea from the sample phase of the microfluidic channel via the diffusion, with a difference of the concentration from the sample flow to both sides of the buffer flow, and an affinity of metal ions into the urea between the buffer phase and the affinity phase. The removal efficiency for the urea was 67, 64, and 63%, with concentrations of 50 mM Mn2+, 10 mM Zn2+, and 5 mM Fe3+ metal ions in the affinity phase, respectively. In addition, protein after desalting with the microfluidic device was improved to more than 10% of the relative activity, with a significant improvement of the signal of mass spectrum shown by MALDI-MS. |
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