Improved agarose gel electrophoresis method and molecular mass calculation for high molecular mass hyaluronan

被引：65

作者：

Cowman, Mary K. ^{[1
]}

Chen, Cherry C. ^{[1
]}

Pandya, Monika ^{[1
]}

Yuan, Han ^{[1
]}

Ramkishun, Dianne ^{[1
]}

LoBello, Jaclyn ^{[1
]}

Bhilocha, Shardul ^{[1
]}

Russell-Puleri, Sparkle ^{[1
]}

Skendaj, Eraldi ^{[1
]}

Mijovic, Jovan ^{[1
]}

Jing, Wei ^{[2
]}

机构：

[1] NYU, Polytech Inst, Brooklyn, NY 11201 USA

[2] Hyalose, Oklahoma City, OK 73104 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2011年 / 417卷 / 01期

关键词：

Hyaluronan; Electrophoresis; Molecular mass; Molecular weight; Solution viscosity; Heat degradation; FACTOR RECEPTOR ACTIVATION; CHEMOKINE GENE-EXPRESSION; SMOOTH-MUSCLE-CELLS; CHEMOENZYMATIC SYNTHESIS; WEIGHT DISTRIBUTION; SODIUM HYALURONATE; FRAGMENTS INDUCE; EPITHELIAL-CELLS; DENDRITIC CELLS; SYNOVIAL-FLUID;

D O I：

10.1016/j.ab.2011.05.023

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

The molecular mass of the polysaccharide hyaluronan (HA) is an important determinant of its biological activity and physicochemical properties. One method currently used for the analysis of the molecular mass distribution of an HA sample is gel electrophoresis. In the current work, an improved agarose gel electrophoresis method for analysis of high molecular mass HA is presented and validated. HA mobility in 0.5% agarose minigels was found to be linearly related to the logarithm of molecular mass in the range from approximately 200 to 6000 kDa. A sample load of 2.5 mu g for polydisperse HA samples was employed. Densitometric scanning of stained gels allowed analysis of the range of molecular masses present in the sample as well as calculation of weight-average and number-average values. The method was validated for a polydisperse HA sample with a weight-average molecular mass of approximately 2000 kDa. Excellent agreement was found between the weight-average molecular mass determined by electrophoresis and that determined by theological measurement of the solution viscosity. The revised method was then used to show that heating solutions of HA at 100 degrees C, followed by various cooling procedures, had no effect on the HA molecular mass distribution. (C) 2011 Elsevier Inc. All rights reserved.

引用

页码：50 / 56

页数：7

共 71 条

[1]

Hyaluronan molecular weight and polydispersity in some commercial intra-articular injectable preparations and in synovial fluid [J].