Partitioning and diffusion of proteins and linear polymers in polyacrylamide gels

被引:138
作者
Tong, J [1 ]
Anderson, JL [1 ]
机构
[1] CARNEGIE MELLON UNIV,DEPT CHEM ENGN,COLLOIDS POLYMERS SURFACES PROGRAM,PITTSBURGH,PA 15213
基金
美国国家科学基金会;
关键词
D O I
10.1016/S0006-3495(96)79712-6
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The equilibrium partition coefficient (K) and diffusion coefficient (D-gel) of two proteins and two linear polymers were measured as a function of polymer content of a 2.7% cross-linked polyacrylamide (PA) gel. The gel concentration, expressed as a volume percentage of PA in the gel (phi), varied between 0 and 14%, The measurements were made by fluorescence spectroscopy; fluorescent dyes were covalently attached to the macromolecules. The dependence of K on phi for the proteins agrees with a model of the gel network as randomly placed, impenetrable rods. The diffusion data are interpreted in terms of an effective medium theory for the mobility of a sphere in a Brinkman fluid, Using values of the Brinkman parameter in the literature, the effective medium model with no adjustable parameters fits the diffusion data for the proteins very well but underpredicts D-gel for the linear polymers. The gel effect on partitioning is significantly greater than that on diffusion. The permeability (KDgel) of bovine serum albumin decreased by 10(3) over the range phi = 0 --> 8%, and the ratio of permeabilities for ribonuclease compared to BSA increased from 2 to 30.
引用
收藏
页码:1505 / 1513
页数:9
相关论文
共 53 条
[1]   PARTICLE DIFFUSION AS A FUNCTION OF CONCENTRATION AND IONIC-STRENGTH [J].
ANDERSON, JL ;
RAUH, F ;
MORALES, A .
JOURNAL OF PHYSICAL CHEMISTRY, 1978, 82 (05) :608-616
[2]  
[Anonymous], RESPONSIVE GELS
[3]   MOBILITY MEASUREMENT BY ANALYSIS OF FLUORESCENCE PHOTOBLEACHING RECOVERY KINETICS [J].
AXELROD, D ;
KOPPEL, DE ;
SCHLESSINGER, J ;
ELSON, E ;
WEBB, WW .
BIOPHYSICAL JOURNAL, 1976, 16 (09) :1055-1069
[4]   ELASTIC PROPERTIES OF HIGHLY CROSS-LINKED POLYACRYLAMIDE GELS [J].
BASELGA, J ;
HERNANDEZFUENTES, I ;
PIEROLA, IF ;
LLORENTE, MA .
MACROMOLECULES, 1987, 20 (12) :3060-3065
[5]   ADVANCED SORBENTS FOR PREPARATIVE PROTEIN SEPARATION PURPOSES [J].
BOSCHETTI, E .
JOURNAL OF CHROMATOGRAPHY A, 1994, 658 (02) :207-236
[6]   PREPARATIVE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATION OF PROTEINS WITH HYPERD ION-EXCHANGE SUPPORTS [J].
BOSCHETTI, E ;
GUERRIER, L ;
GIROT, P ;
HORVATH, J .
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS, 1995, 664 (01) :225-231
[7]  
BRINKMAN HC, 1947, APPL SCI RES, V1, P27
[8]   COMMENTS ON EXCLUSION OF POLYMER-CHAINS FROM SMALL PORES AND ITS RELATION TO GEL-PERMEATION CHROMATOGRAPHY [J].
CASASSA, EF .
MACROMOLECULES, 1976, 9 (01) :182-185
[9]   DIRECT MEASUREMENT OF INTERSTITIAL CONVECTION AND DIFFUSION OF ALBUMIN IN NORMAL AND NEOPLASTIC TISSUES BY FLUORESCENCE PHOTOBLEACHING [J].
CHARY, SR ;
JAIN, RK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (14) :5385-5389
[10]   POLYMERIZATION OF POLYACRYLAMIDE GELS - EFFICIENCY AND REPRODUCIBILITY AS A FUNCTION OF CATALYST CONCENTRATIONS [J].
CHRAMBACH, A ;
RODBARD, D .
SEPARATION SCIENCE, 1972, 7 (06) :663-703