Plasma protein adsorption patterns on emulsions for parenteral administration:: Establishment of a protocol for two-dimensional polyacrylamide electrophoresis

被引:30
作者
Harnisch, S [1 ]
Müller, RH [1 ]
机构
[1] Free Univ Berlin, Dept Pharmaceut Biopharmaceut & Biotechnol, D-12169 Berlin, Germany
关键词
two-dimensional polyacrylamide gel electrophoresis; immobilized pH gradients; thiourea; apolipoproteins; fat emulsions; particle size;
D O I
10.1002/elps.1150190233
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The two-dimensional polyacrylamide electrophoresis (2-D PAGE) of the plasma protein adsorption pattern previously established for polymeric nanoparticles was modified and transferred to oil in water emulsions for intravenous administration. The emulsions were incubated with citrated plasma, and separation from excess plasma was performed by centrifugation under optimized conditions: 15 000 g and three washing steps with 0.05 M phosphate buffer, pH 7.4. With this sample preparation, coalescence of droplets could be avoided and an unchanged surface area maintained, in addition the phosphate buffer minimized artificial IgG adsorption. Critical factors affecting sensitivity were contamination of the sample by oil residues and the use of thiourea in the immobilized pH gradients. Changes in the protein adsorption pattern caused by altered surface properties of the emulsion (i.e. adsorbed Poloxamer 407) were detectable when applying the optimized protocol. Knowledge of the protein adsorption patterns and their correlation to in vivo behavior opens the perspective for the development of intravenous emulsions for controlled drug delivery.
引用
收藏
页码:349 / 354
页数:6
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