Modification of microfiltration membranes by hydrogel impregnation for pDNA purification

被引:12
作者
Castilho, Pedro H. [1 ]
Correia, Tiago R. [1 ]
de Amorim, Maria T. Pessoa [2 ]
Escobar, Isabel C. [2 ,3 ]
Queiroz, Joao A. [1 ]
Correia, Ilidio J. [1 ]
Morao, Antonio M. [1 ]
机构
[1] Univ Beira Interior, Fac Hlth Sci, Hlth Sci Res Ctr, CICS UBI, Covilha, Portugal
[2] Univ Minho, Dept Text Engn, P-4800058 Guimaraes, Portugal
[3] Univ Toledo, Dept Chem & Environm Engn, Toledo, OH 43606 USA
关键词
membranes; nucleic acids; polysaccharides; SUPERCOILED PLASMID DNA; SURFACE MODIFICATION; ULTRAFILTRATION; DIFFUSION; TRANSPORT; MODEL; TRANSMISSION; SEPARATION; RECOVERY;
D O I
10.1002/app.41610
中图分类号
O63 [高分子化学(高聚物)];
学科分类号
070305 [高分子化学与物理];
摘要
The huge efforts for the achievement of highly purified biomolecules are growing every day. A great number of efficient techniques, such as chromatography, are already available in laboratory for separation processes. However, membrane-based technologies are the best match to assure simplicity, efficiency and easy scale-up procedures. Herein we report the modification of a commercial microfiltration membrane for plasmid DNA purification by agarose gel impregnation. The membrane was characterized by SEM, ATR-FTIR, EDS, contact angle, and porosity measurements. Additionally, the membrane pore radius was estimated from observed rejections of different proteins and with that information the rejection of a 6050 bp plasmid DNA (pDNA) molecule was estimated for different values of flux using a theoretical model of large flexible molecules in membranes with parallel cylindrical pores, which is applicable to pDNA ultrafiltration in conventional membranes, as recently shown in the literature. The experimental results show that the modified membrane has higher pDNA rejections than the predicted by the model, suggesting that the different type of porous structure that a hydrogel has, may have a positive effect on pDNA rejections as compared to other biomolecules with more rigid structures, making this type of modified membranes potential better candidates to be used for the selective recovery of pDNA in this type of bioprocesses. (c) 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41610.
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页数:8
相关论文
共 35 条
[1]
Supermacroprous chitosan-agarose-gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering [J].
Bhat, Sumrita ;
Tripathi, Anuj ;
Kumar, Ashok .
JOURNAL OF THE ROYAL SOCIETY INTERFACE, 2011, 8 (57) :540-554
[2]
A bi-layer electrospun nanofiber membrane for plasmid DNA recovery from fermentation broths [J].
Correia, Tiago R. ;
Antunes, Bernardo P. ;
Castilho, Pedro H. ;
Nunes, Jose C. ;
Pessoa de Amorim, Maria T. ;
Escobar, Isabel C. ;
Queiroz, Joao A. ;
Correia, Ilidio J. ;
Morao, Antonio M. .
SEPARATION AND PURIFICATION TECHNOLOGY, 2013, 112 :20-25
[3]
HINDERED TRANSPORT OF LARGE MOLECULES IN LIQUID-FILLED PORES [J].
DEEN, WM .
AICHE JOURNAL, 1987, 33 (09) :1409-1425
[4]
Fuji T, 2009, TISSUE ENG PT A, V15, P3525, DOI [10.1089/ten.tea.2009.0048, 10.1089/ten.TEA.2009.0048]
[5]
Formulation of chitosan-TPP-pDNA nanocapsules for gene therapy applications [J].
Gaspar, V. M. ;
Sousa, F. ;
Queiroz, J. A. ;
Correia, I. J. .
NANOTECHNOLOGY, 2011, 22 (01)
[6]
Current trends in separation of plasmid DNA vaccines: A review [J].
Ghanem, Ashraf ;
Healey, Robert ;
Adly, Frady G. .
ANALYTICA CHIMICA ACTA, 2013, 760 :1-15
[7]
Thermosensitive sol-gel reversible hydrogels [J].
Jeong, B ;
Kim, SW ;
Bae, YH .
ADVANCED DRUG DELIVERY REVIEWS, 2002, 54 (01) :37-51
[8]
Hindered diffusion in agarose gels: Test of effective medium model [J].
Johnson, EM ;
Berk, DA ;
Jain, RK ;
Deen, WM .
BIOPHYSICAL JOURNAL, 1996, 70 (02) :1017-1023
[9]
Kahn DW, 2000, BIOTECHNOL BIOENG, V69, P101, DOI 10.1002/(SICI)1097-0290(20000705)69:1<101::AID-BIT12>3.0.CO
[10]
2-1