SOLID-PHASE IMMUNE-REACTIONS AS MONITORED BY SEDIMENTATION FIELD-FLOW FRACTIONATION

被引:15
作者
LANGWOST, B [1 ]
CALDWELL, KD [1 ]
机构
[1] UNIV UTAH,DEPT BIOENGN,SALT LAKE CITY,UT 84112
关键词
FIELD-FLOW FRACTIONATION; IMMUNE COMPLEX; ANTIBODY COATING; LAT; PROTEIN ADSORPTION;
D O I
10.1007/BF02268362
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Sedimentation field-flow fractionation was shown to permit the precise evaluation of surface concentrations of human IgG, adsorbed to polystyrene latex spheres of different sizes. Unlike conventional techniques for measuring protein uptake by colloidal substrates, this method allowed a direct evaluation of mass adsorbed per unit area, without the need for potentially destructive labelling reactions. Thus, a four hour adsorption of IgG from a 3-10 fold excess of protein in solution yielded surface concentrations which were 1.4 +/- 0.1 mg/m2 on a 272 nm latex and 1.9 +/- 0.1 mg/m2 on a latex with a diameter of 142 nm. The lower value coincided with the estimated monolayer surface coverage. The IgG-PS 272 nm adsorption complex was shown to take up negligible amounts of HSA from a 10 mg/mL solution, while its specific uptake of a polyclonal rabbit antihuman IgG was 2.6 molecules per molecule of adsorbed antigen. The same ratio was found for the smaller particles. The surface concentration of adsorbed second antibody, often crucial in immunodiagnostic quantifications, was therefore found to be significantly enhanced by the increased substrate curvature presented by the smaller particles.
引用
收藏
页码:317 / 324
页数:8
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