Ivanov Alexander E, Galaev Igor Yu, Mattiasson Bo
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, SE-221 00 Lund, Sweden.
J Mol Recognit. 2006 Jul-Aug;19(4):322-31. doi: 10.1002/jmr.792.
Interaction of mono- and disaccharides, polysaccharide particles and yeast cells with boronate-containing copolymers (BCC) of N-acryloyl-m-aminophenylboronic acid (NAAPBA) with N,N-dimethylacrylamide (DMAA) or N-isopropylacrylamide (NIPAM) was studied. The binding of saccharides to BCC of NIPAM resulted in a shift of its phase transition temperature (DeltaTP), which provided a quantitative measure for the complex formation. Among the sugars typical of non-reducing ends of glycoproteins the DeltaTP decreased in the order: N-acetylneuraminic acid > xylose approximately galactose > mannose approximately fucose >> N-acetylglucosamine. Strong specific adsorption of the BCC on the cross-linked agarose gel Sepharose CL-6B (15-30 mg/ml gel at pH 9.2) was registered. The copolymers adsorption was due to boronate-sugar interactions and decreased with pH. Multivalent interaction of the BCC with the agarose gel has been proven by liquid column chromatography exhibiting a weak reversible adsorption of NAAPBA and almost irreversible adsorption of DMAA-NAAPBA copolymer from 0.1 M sodium phosphate buffer, pH 7.9. The two studied BCCs could be completely desorbed from the gel by 0.1 M fructose in aqueous buffered media with pH from 7.5 to 9.2. In turn, the agarose particles and yeast cells were found to adhere to siliceous supports end-grafted with boronate-BCC of N,N-dimethylacrylamide at pH > or = 7.5, due to the actions. Quantitative detachment of adhered particles or cells could be attained by addition of 20 mM or 100 mM fructose, respectively, in the pH range from 7.5 to 9.2. Affinity adhesion of micron-size carbohydrate particles to boronate-containing polymer brushes fixed on solid supports was considered as a model system suggesting a new approach to isolation and separation of living cells.
研究了单糖和双糖、多糖颗粒以及酵母细胞与含硼酸酯的N-丙烯酰基间氨基苯硼酸(NAAPBA)与N,N-二甲基丙烯酰胺(DMAA)或N-异丙基丙烯酰胺(NIPAM)的共聚物(BCC)之间的相互作用。糖类与NIPAM的BCC结合导致其相变温度(ΔTP)发生变化,这为复合物的形成提供了一种定量测量方法。在糖蛋白非还原端典型的糖类中,ΔTP按以下顺序降低:N-乙酰神经氨酸>木糖≈半乳糖>甘露糖≈岩藻糖>>N-乙酰葡糖胺。记录到BCC在交联琼脂糖凝胶Sepharose CL-6B上有强烈的特异性吸附(在pH 9.2时为15 - 30 mg/ml凝胶)。共聚物的吸附是由于硼酸酯 - 糖相互作用,且随pH降低。通过液相柱色谱法证明了BCC与琼脂糖凝胶的多价相互作用,该方法显示从pH 7.9的0.1 M磷酸钠缓冲液中,NAAPBA有弱可逆吸附,而DMAA - NAAPBA共聚物几乎不可逆吸附。在pH为7.5至9.2的水性缓冲介质中,0.1 M果糖可使两种研究的BCC从凝胶上完全解吸。反过来,发现琼脂糖颗粒和酵母细胞在pH≥7.5时由于这些作用而粘附到末端接枝有N,N-二甲基丙烯酰胺硼酸酯 - BCC的硅质载体上。在pH 7.5至9.2范围内,分别加入20 mM或100 mM果糖可实现粘附颗粒或细胞的定量脱离。微米级碳水化合物颗粒与固定在固体载体上的含硼酸酯聚合物刷的亲和粘附被视为一种模型系统,为活细胞的分离和分离提出了一种新方法。