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使用重量法压电石英生物传感器测定脂多糖与壳聚糖及其N-乙酰化衍生物之间的结合参数。

Determination of the parameters of binding between lipopolysaccharide and chitosan and its N-acetylated derivative using a gravimetric piezoquartz biosensor.

作者信息

Naberezhnykh G A, Gorbach V I, Kalmykova E N, Solov'eva T F

机构信息

G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia; Far East Federal University, Vladivostok, Russia.

G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far East Branch of Russian Academy of Sciences, Vladivostok, Russia.

出版信息

Biophys Chem. 2015 Mar;198:9-13. doi: 10.1016/j.bpc.2015.01.003. Epub 2015 Jan 10.

Abstract

The interaction of endotoxin (lipopolysaccharide - LPS) with low molecular weight chitosan (5.5 kDa), its N-acylated derivative and chitoliposomes was studied using a gravimetric piezoelectric quartz crystal microbalance biosensor. The optimal conditions for the formation of a biolayer based on immobilized LPS on the resonator surface and its regeneration were elaborated. The association and dissociation rate constants for LPS binding to chitosans were determined and the affinity constants (Kaf) were calculated based on the data on changes in the oscillation frequency of the quartz crystal resonator. The Kaf values correlated with the ones obtained using other methods. The affinity of N-acylated chitosan binding to LPS was higher than that of the parent chitosan binding to LPS. Based on the results obtained, we suggest that water-soluble N-acylated derivatives of chitosan with low degree of substitution of amino groups could be useful compounds for endotoxin binding and neutralization.

摘要

使用重量法压电石英晶体微天平生物传感器研究了内毒素(脂多糖-LPS)与低分子量壳聚糖(5.5 kDa)、其N-酰化衍生物和壳聚糖脂质体的相互作用。阐述了基于固定在谐振器表面的LPS形成生物层及其再生的最佳条件。测定了LPS与壳聚糖结合的缔合和解离速率常数,并根据石英晶体谐振器振荡频率变化的数据计算了亲和常数(Kaf)。Kaf值与使用其他方法获得的值相关。N-酰化壳聚糖与LPS结合的亲和力高于母体壳聚糖与LPS结合的亲和力。基于所得结果,我们认为氨基取代度低的水溶性N-酰化壳聚糖衍生物可能是用于内毒素结合和中和的有用化合物。

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