Complement activation by model drug carriers for intravenous application: determination by two-dimensional electrophoresis.
The interactions of intravenously injected drug carriers with blood proteins are considered as an important factor for the fate of the particles after their administration. Protein adsorption on latex particles applied as model for intravenous drug carriers was analysed using two-dimensional electrophoresis (2-DE). The particles were incubated in citrated plasma, serum and heat-inactivated serum, respectively. Incubation in the various media resulted in clear differences in the protein adsorption patterns. Two characteristic protein spots were determined to be enriched on the 2-DE gels only after incubation of the particles in serum. Employing N-terminal microsequencing these protein spots were identified to be fragments of the complement protein C3. Enrichment of these particular spots was most likely a result of complement activation by the particles. Mechanism of C3 binding to the particle surface and subsequent inactivation by cleavage are discussed in order to explain the results. It could be demonstrated that 2-DE analysis provides the possibility to distinguish between adsorption and covalent attachment of C3 to particulate surfaces. The findings indicate that complement activation was caused by covalent binding of the C3 component C3b to the particles' surface. The influence of the incubation medium on the in vitro protein adsorption of particulate drug carriers has to be considered when a correlation between the protein adsorption pattern and the in vivo behaviour of the particles is approached.
Lück
M
M
Department of Pharmaceutics, Biopharmaceutics and Biotechnology, The Free University of Berlin, Germany.
Schröder
W
W
Paulke
B R
BR
Blunk
T
T
Müller
R H
RH
D000327:Adsorption; D003167:Complement Activation / Q000187:drug effects*; D003176:Complement C3 / Q000737:chemistry / Q000378:metabolism*; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007275:Injections, Intravenous; D007840:Latex / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008956:Models, Chemical; D013499:Surface Properties
D003176:Complement C3; D004337:Drug Carriers; D007840:Latex