Adsorption and recognition of protein molecular imprinted calcium alginate/polyacrylamide hydrogel film with good regeneration performance and high toughnessby Kongyin Zhao, Tian Chen, Beibei Lin, Wenkui Cui, Bohong Kan, Ning Yang, Xiangyu Zhou, Xinxin Zhang, Junfu Wei

Reactive and Functional Polymers

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School of Material Science and Engineering, Tianjin Polytechnic University, Tianjin 300387, China c The First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China a r t i c l e i n f o

Article history: cient way to prepare synthetic materials bearing selective molecusors [3], drug release [4] and environmental protection [5,6]. The imprinting of low-molecular weight compound has been well established. Nevertheless, several challenges remain in the imprinting of bio-macromolecules, such as proteins, DNAs, and for the molecular imprinting of proteins [10–12]. PAM hydrogel is us structure. The m strong interacaqueous system ers have a lly induce radical polymerization [14–22]. However, the PAM hydrog low cross-linking degree and poor mechanical properties results in low affinity and poor regeneration properties template [23]. The high cross-linked PAM hydrogel effectively depresses the swelling of gel and the deformation of imprinting cavities, improving the affinity of the polymer. Nevertheless, as the cross-linking density increases, the transfer of protein was restricted [24]. Addressing these problems is usually attempted by adding another component to improve the mechanical performance and decrease the swelling of the hydrogel [11–13]. ⇑ Corresponding author at: State Key Laboratory of Hollow Fiber Membrane

Materials and Processes, Tianjin Polytechnic University, Tianjin 300387, China. Tel.: +86 02283955362; fax, +86 0228395055.

E-mail address: zhaokongyin@tjpu.edu.cn (K. Zhao).

Reactive & Functional Polymers 87 (2015) 7–14

Contents lists availab

Reactive & Funct .elar recognition sites and binding pockets at the surface or inside polymer matrix. Molecularly imprinted polymers (MIPs) are characterized by their thermal and chemical stability, high specificity, ease of mass preparation, low cost and reusability, which promote their wide applications in chromatography [1], catalysis [2], senbiocompatible and has soft and wet macroporo abundant amide functional groups in PAM can for tions with peptide bonds in the proteins even in [13]. Protein imprinted acrylamide-based polym been successfully fabricated by UV or thermahttp://dx.doi.org/10.1016/j.reactfunctpolym.2014.12.001 1381-5148/ 2014 Elsevier B.V. All rights reserved.lready d free el with , often for thewithout residue eluent should be used to prepare MIP for cell culture.  2014 Elsevier B.V. All rights reserved. 1. Introduction

Molecule imprinting technology has been proved to be an effieven whole cells and viruses, due to their large size, structural complexity and the flexible conformation [7–9].

Polyacrylamide (PAM) hydrogel was widely used as the matrixReceived 24 November 2014

Accepted 15 December 2014

Available online 22 December 2014

Keywords:

Protein molecular imprinting

Calcium alginate/polyacrylamide

Hydrogel film

Adsorption

Recognition

Cell culturea b s t r a c t

Protein imprinted calcium alginate/polyacrylamide hydrogel film (CA/PAMMIP) with high toughness was prepared using bovine serum albumin (BSA) as template molecule, sodium alginate and acrylamide as functional monomers, N,N0-methylenebisacrylamide (MBAA) as the covalent cross-linker and CaCl2 as the ionic cross-linker via UV radiation-reduced polymerization. Factors affecting the adsorption capacity and imprinting efficiency of the BSA-imprinted CA/PAM hydrogel films were investigated, such as ratio of polyacrylamide/sodium alginate, film thickness, MBAA concentration and CaCl2 concentration.

Results showed that the CA/PAM MIP exhibited an obvious improvement in terms of adsorption capacity for BSA compared with non-imprinted polymer (NIP). The adsorption capacity of MIP for BSA reached 22.49 mg/g, which was 2.7 times higher than NIP. The regeneration property of the BSA-imprinted CA/

PAM hydrogel was distinctly improved and the imprinting efficiency of CA/PAM MIP maintained 77.95% of the initial value after five repetitions. Single and binary proteins rebinding indicated that the

CA/PAM MIP exhibited good recognition performance. Cell culture experiments showed CA/PAM MIP was more suitable for cell culture than CA/PAM NIP. The residual sodium dodecyl sulfate (SDS) in the elution process leaded to the death of mouse fibroblast cells (L929) after 3 days. A moderate elution solutiona State Key Laboratory of Hollow Fiber Membrane Materials and Processes, Tianjin Polytechnic University, Tianjin 300387, China bAdsorption and recognition of protein m alginate/polyacrylamide hydrogel film w performance and high toughness

Kongyin Zhao a,b,⇑, Tian Chen b, Beibei Lin b, Wenkui

Xinxin Zhang b, Junfu Wei a,b journal homepage: wwwcular imprinted calcium good regeneration i b, Bohong Kan c, Ning Yang b, Xiangyu Zhou a, le at ScienceDirect ional Polymers lsev ier .com/ locate/ react exhibit good performance in maintaining the imprinted cavities dodecyl sulfate (SDS) left in the hydrogel, which leaded to the ctiondeath of L929 cells after 3 days. 2. Experimental 2.1. Materials

Acrylamide (AM), N,N0-methylenebisacrylamide (MBAA) and ammoniumpersulfate (APS) were purchased from Kermel Chemical (Tianjin, China). Sodium dodecyl sulfate (SDS) and glacial acetic acid (HAc) were purchased from Institute of Guangfu Fine Chemicals (Tianjin, China). Sodium alginate (SA) was obtained from Sinopharm Chemical Reagent co., Ltd. Bovine serum albumin (BSA), ovalbumin (Ova), bovine hemoglobin (Hb) and bovine c-globulin (Glo) were purchased from Shanghai Science and Technology

Development Company (Shanghai, China). Calcium chloride dehydrate was supplied by Yingda Chemicals (Tianjin, China). Mouse fibroblast cells (L929) were obtained from the Cellular Biology

Institute of the Chinese Academy of Sciences (Shanghai, China). 2.2. Preparation of CA/PAM MIP

Fig. 1 shows the schematic representation for the fabrication of

CA/PAM MIP. Unless otherwise stated, the water content was fixed at approximate 86 wt%. The mass ratio of AM to SA was 6:1, 8:1, 10:1 and 12:1, respectively. The weight of MBAA and APS was fixed at 0.09and 1% of acrylamide, respectively. AM, MBAA, SA, APS and