Determination of concentration and aggregate size in influenza virus preparations from true UV absorption spectra

被引：0

作者：

Ksenofontov A.L. ^{[1
]}

Kozlovskii V.S. ^{[1
]}

Kordyukova L.V. ^{[1
]}

Radyukhin V.A. ^{[1
]}

Timofeeva A.V. ^{[1
]}

Dobrov E.N. ^{[1
]}

机构：

[1] Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow

来源：

Molecular Biology | 2006年 / 40卷 / 1期

基金：

俄罗斯基础研究基金会;

关键词：

Extrapolation method; Influenza virus; Light scattering; UV spectra; Virion aggregation;

D O I：

10.1134/S0026893306010201

中图分类号：

学科分类号：

摘要：

Light scattering is known to make a considerable contribution to ultraviolet absorption spectra of influenza virus (Flu) preparations. We applied extrapolation to analysis of this contribution. Ultraviolet spectra were recorded and true extinction coefficients (A1 cm, 2800.1%) were determined in suspensions of intact virions of Flu strain Puerto Rico/8/34 and subviral particles obtained by bromelain digestion of the same strain (1.26 ± 0.17 and 0.96 ± 0.11 OD, respectively). This allowed simple and rapid measurement of virus concentration. It was shown that UV spectra allowed efficient monitoring of virion aggregation. The pH dependence of aggregation properties of influenza subviral particles was studied. © Pleiades Publishing, Inc., 2006.

引用

页码：152 / 158

页数：6

共 18 条

[1]

Gill S.C., Von Hippel P.H., Calculation of protein extinction coefficients from amino acid sequence data, Anal. Biochem., 182, pp. 319-326, (1989)

[2]

Tikchonenko T.I., Dobrov E.N., Velikodvorskaya G.A., Kisseleva N.P., Peculiarities of the secondary structure of phage DNA in situ, J. Mol Biol., 18, pp. 58-67, (1966)

[3]

Yokote Y., Arai K.M., Akahane K., Recovery of tryptophan from 25-minute acid hydrolysates of protein, Anal. Biochem., 152, pp. 245-249, (1986)

[4]

Laemmli U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227, pp. 680-685, (1970)

[5]

Peterson G.L., A simplification of the protein assay method of Lowry et al. which is more generally applicable, Anal. Biochem., 83, pp. 346-356, (1977)

[6]

Brand C.M., Skehel J.J., Crystalline antigen from the influenza virus envelope, Nature New. Biol., 238, pp. 145-147, (1972)

[7]

Chu B., Laser light scattering, Ann. Rev. Phys. Chem., 21, pp. 145-174, (1970)

[8]

Kulburn E.D., Influenza Viruses and Influenza [Russian Translation], (1978)

[9]

Camerini-Otero R.D., Franklin R.M., Day L.A., Molecular weights, dispersion of refractive index increments, and dimensions from transmittance spectrophotometry. Bacteriophages R17, T7, and PM2, and tobacco mosaic virus, Biochemistry, 13, pp. 3763-3773, (1974)

[10]

Camerini-Otero R.D., Day L.A., The wavelength dependence of the turbidity of solutions of macromolecules, Biopolymers, 17, pp. 2241-2249, (1978)

← 1 2 →