Thermoresponsive Poly（N，N-dimethylacrylamide-co-N-isopropylacrylamide） for Preparation of Composite Sieving Media in Non-Gel Capillary Electrophoresis

doi:10.19894/j.issn.1000-0518.230125

Chinese Journal of Applied Chemistry ›› 2023, Vol. 40 ›› Issue (9): 1312-1321.DOI: 10.19894/j.issn.1000-0518.230125

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Thermoresponsive Poly（N，N-dimethylacrylamide-co-N-isopropylacrylamide） for Preparation of Composite Sieving Media in Non-Gel Capillary Electrophoresis

Feng-Hao JIANG¹^,², Han HONG³, Bo-Wei JIANG⁴, Zhao-Hui SU¹^,²()

^1.State Key Laboratory of Polymer Physics and Chemistry，Changchun Institute of Applied Chemistry，Chinese Academy of Sciences，Changchun 130022，China
^2.University of Science and Technology of China，Hefei 230026，China
^3.Lifepolymer Composite Material Technology Co. ，Ltd. ，Taizhou 225321，China
^4.The First Research Institute of the Ministry of Public Security，Beijing 100048，China

Received:2023-04-26 Accepted:2023-07-27 Published:2023-09-01 Online:2023-09-14
Contact: Zhao-Hui SU
About author:zhsu@ciac.ac.cn

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Abstract

Abstract:

In this paper， a random copolymer of N，N-dimethylacrylamide and N-isopropylacrylamide （P（DMA-co-NIPAM）） as well as poly-N?，?N-dimethylacrylamide （PDMA） are synthesized and blended to prepare a composite sieving medium for non-gel capillary electrophoresis， aiming to reduce the viscosity of the sieving medium while enhance its DNA separation performance. Analyses by NMR spectroscopy and Fourier transform infrared （FT-IR） spectroscopy confirm that both polymers are successfully synthesized. The hydrodynamic diameter， low shear viscosity and UV-Vis absorbance results show that P（DMA-co-NIPAM） is thermoresponsive with a low critical solution temperature （LCST） of 60~80 ℃. DNA sieving results indicate that the composite medium containing 10% P（DMA-co-NIPAM） exhibits best properties， a viscosity 19% lower than PDMA and a higher resolution and theoretical plate number in separating large DNA fragments， and separation time is reduced by more than 4%， showing good separation performance.

Key words: Capillary electrophoresis, Non-gel sieving media, DNA separation, Thermoresponsive polymers, Poly（N, N-dimethylacrylamide）

CLC Number:

O631

Feng-Hao JIANG, Han HONG, Bo-Wei JIANG, Zhao-Hui SU. Thermoresponsive Poly（N，N-dimethylacrylamide-co-N-isopropylacrylamide） for Preparation of Composite Sieving Media in Non-Gel Capillary Electrophoresis[J]. Chinese Journal of Applied Chemistry, 2023, 40(9): 1312-1321.

Figures/Tables 9

Fig.1 Schematic diagram of the transition process of P（DMA-co-NIPAM） with increasing temperature

Table 1 Composition of the composite NGCE sieving media

NGCE sieving media	n（P（DMA-co-NIPAM））∶ n（PDMA）	w（polymer）/%	c（Urea）/（mol·L^-1）	w（2-pyrrolidone）/%
PDMA ^a	0∶10	4	8	5.7
Ⅰ	1∶9
Ⅱ	2∶8
Ⅲ	5∶5
P（DMA-co-NIPAM） ^b	10∶0

Fig.2 Synthesis route of PDMA （A） and P（DMA-co-NIPAM）（B）

Fig.3 Temperature dependence of the hydrodynamic diameter （A）， viscosity （B） and absorbance （C） of polymer solutions with different ratios of P （DMA-co-NIPAM） to PDMA. The mass concentrations of the solutions are all 20 mg/mL

Fig.4 Viscosities of five NGCE sieving media at 25 ℃

Fig.5 600 LIZ DNA capillary electrophoresis separation patterns using NGCE media PDMA，Ⅰ，Ⅱ and Ⅲ

Fig.6 The resolution （A）， plate numbers （B）， electrophoresis completion time （C）， and linearity of migration time versus number of nucleotides （D） for the composite NGCE sieving media in separating standard DNA samples at 60 ℃

Table 2 Linear regression coefficient of a plot of DNA fragment migration time versus nucleotides number

Sample	PDMA	Ⅰ	Ⅱ	Ⅲ
Linear regression coefficient（R²）	0.997 0	0.996 2	0.994 8	0.989 0

Fig.7 The resolution （A） and plate numbers （B） for the composite NGCE sieving media in separating standard DNA samples （414/420 bp） at different temperatures

References 20

1	陈义. 毛细管电泳技术及应用［M］. 北京：化学工业出版社， 2006： 4-5.
	CHEN Y. Capillary electrophoresis techniques and applications［M］. Beijing： Chemical Industry Press， 2006： 4-5.
2	SARTORI A， BARBIER V， VIOVY J L. Sieving mechanisms in polymeric matrices［J］. Electrophoresis， 2003， 24（3）： 421-440.
3	HELLER C. Capillary electrophoresis of proteins and nucleic acids in gels and entangled polymer solutions［J］. J Chromatogr A， 1995， 698（1/2）： 19-31.
4	ASHTON R， PADALA C， KANE R S. Microfluidic separation of DNA［J］. Curr Opin Biotechnol， 2003， 14（5）： 497-504.
5	王前，许旭. 用于毛细管电泳DNA分离的合成聚合物［J］. 化学进展， 2003（4）： 275-287.
	WANG Q， XU X. Synthetic polymers for DNA separation by capillary electrophoresis［J］. Prog Chem， 2003（4）： 275-287.
6	HEIGER D N， COHEN A S， KARGER B L. Separation of DNA restriction fragments by high performance capillary electrophoresis with low and zero crosslinked polyacrylamide using continuous and pulsed electric fields［J］. J Chromatogr A， 1990， 516（1）： 33-48.
7	CHIARI M， MICHELETTI C， NESI M， et al. Towards new formulations for polyacrylamide matrices： N-acryloylaminoethoxyethanol， a novel monomer combining high hydrophilicity with extreme hydrolytic stability［J］. Electrophoresis， 1994， 15（2）： 177-186.
8	GAO Q， YEUNG E S. A matrix for DNA separation： genotyping and sequencing using poly（vinylpyrrolidone） solution in uncoated capillaries［J］. Anal Chem， 1998， 70（7）： 1382-1388.
9	KIM Y， MORRIS M D. Separation of nucleic acids by capillary electrophoresis in cellulose solutions with mono- and bis-intercalating dyes［J］. Anal Chem， 1994， 66（7）： 1168.
10	KIM Y， YEUNG E S. Separation of DNA sequencing fragments up to 1000 bases by using poly（ethylene oxide）-filled capillary electrophoresis［J］. J Chromatogr A， 1997， 781（1/2）： 315-325.
11	CHIARI M， MELIS A. Low viscosity DNA sieving matrices for capillary electrophoresis［J］. TrAC Trends Anal Chem， 1998， 17（10）： 623-632.
12	CHIHFIELD C L， HOLLAND L A. Protein sieving with capillary nanogel electrophoresis［J］. Anal Chem， 2021， 93（3）： 1537-1543.
13	SUDOR J， BARBIER V， THIROT S， et al. New block-copolymer thermoassociating matrices for DNA sequencing： effect of molecular structure on rheology and resolution［J］. Electrophoresis， 2001， 22（4）： 720-728.
14	BARKER I C， COWIE J M G， HUCKERBY T N， et al. Studies of the “smart” thermoresponsive behavior of copolymers of N-isopropylacrylamide and N，N-dimethylacrylamide in dilute aqueous solution［J］. Macromolecules， 2003， 36（20）： 7765-7770.
15	ZHANG Y， WANG P， CHEN R. Effect of the macromolecular architecture on the thermoresponsive behavior of poly（N-isopropylacrylamide） in copolymers with poly（N，N-dimethylacrylamide） in aqueous solutions： block vs random copolymers［J］. React Funct Polym， 2022， 171： 105150.
16	ROSENBLUM B B， OAKS F， MENCHEN S， et al. Improved single-strand DNA sizing accuracy in capillary electrophoresis［J］. Nucleic Acids Res， 1997， 25（19）： 3925-3929.
17	CHEN S， WANG K， ZHANG W. A new thermoresponsive polymer of poly（N-acryloylsarcosine methyl ester） with a tunable LCST［J］. Polym Chem， 2017， 8（20）： 3090-3101.
18	BOULOS S， CABRICES O， BLAS M， et al. Development of an entangled polymer solution for improved resolution in DNA typing by CE［J］. Electrophoresis， 2008， 29（23）： 4695-4703.
19	何曼君. 高分子物理［M］. 上海：复旦大学出版社， 2008： 142-144.
	HE M J. Polymer physics［M］. Shanghai： Fudan University Press， 2008： 142-144.
20	LIU R， DE LEONARDIS P， CELLESI F， et al. Cationic temperature-responsive poly（N-isopropyl acrylamide） graft copolymers： from triggered association to gelation［J］. Langmuir， 2008， 24（14）： 7099-7106.

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Thermoresponsive Poly（N，N-dimethylacrylamide-co-N-isopropylacrylamide） for Preparation of Composite Sieving Media in Non-Gel Capillary Electrophoresis

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