Ivanov Alexander E, Ljunggren Lennart
VitroSorb AB, Medeon Science Park, Per Albin Hanssons Väg 41, SE-20512, Malmö, Sweden.
Malmö University, Faculty of Health and Society, Department of Biomedical Science, SE-20506, Malmö, Sweden.
Heliyon. 2019 Nov 27;5(11):e02913. doi: 10.1016/j.heliyon.2019.e02913. eCollection 2019 Nov.
Thin macroporous poly(vinyl alcohol) (PVA) hydrogels were produced by cross-linking of PVA in a semi-frozen state with glutaraldehyde (GA) on glass slides or in the wells of microtiter plates. The 100-130 μm-thick gels were mechanically transferable, squamous translucent films with a high porosity of 7.2 ± 0.3 mL/g dry PVA similar to larger cylindrical PVA monoliths of the same composition. Additional treatment of the gels with 1% GA increased the aldehyde group content from 0.7 to 2.4 μmol/mL as estimated using dinitrophenylhydrazine (DNPH) reagent. Translucency of the gels allowed registration of UV-visible spectra of the DNPH-stained films. The catalytic activity of trypsin covalently immobilized on thin gels in the microtiter plates was estimated with chromogenic substrate directly in the wells, and indicated that the amount of protein immobilized was at least 0.34 mg/mL gel. Human immunoglobulin G (IgG) immobilized on thin gels at 0.1-10 mg/mL starting concentrations could be detected in a concentration-dependent manner due to recognition by anti-human rabbit IgG conjugated with peroxidase and photometric registration of the enzymatic activity. The results indicate good permeability of the hydrogel pores for macromolecular biospecific reagents and suggest applications of thin reactive PVA hydrogels in photometric analytical techniques.
通过在半冷冻状态下用戊二醛（GA）使聚乙烯醇（PVA）在载玻片或微量滴定板孔中交联，制备了薄的大孔PVA水凝胶。这种100 - 130μm厚的凝胶是可机械转移的鳞状半透明薄膜，具有7.2±0.3 mL/g干PVA的高孔隙率，类似于相同组成的较大圆柱形PVA整块材料。用1% GA对凝胶进行额外处理后，使用2,4 - 二硝基苯肼（DNPH）试剂估计醛基含量从0.7增加到2.4μmol/mL。凝胶的半透明性使得可以记录DNPH染色薄膜的紫外可见光谱。直接在微量滴定板孔中用显色底物估计共价固定在薄凝胶上的胰蛋白酶的催化活性，结果表明固定的蛋白量至少为0.34 mg/mL凝胶。起始浓度为0.1 - 10 mg/mL固定在薄凝胶上的人免疫球蛋白G（IgG），由于与过氧化物酶偶联的抗人兔IgG的识别以及酶活性的光度记录，可以以浓度依赖的方式检测到。结果表明水凝胶孔对大分子生物特异性试剂具有良好的渗透性，并表明薄的反应性PVA水凝胶在光度分析技术中的应用前景。
