M. Luck et al., Complement activation by model drug carriers for intravenous application: determination by two-dimensional electrophoresis, BIOMATERIAL, 20(21), 1999, pp. 2063-2068
The interactions of intravenously injected drug carriers with blood protein
s are considered as an important factor for the fate of the particles after
their administration. Protein adsorption on latex particles applied as mod
el for intravenous drug carriers was analysed using two-dimensional electro
phoresis (2-DE). The particles were incubated in citrated plasma, serum and
heat-inactivated serum, respectively. Incubation in the various media resu
lted in clear differences in the protein adsorption patterns. Two character
istic protein spots were determined to be enriched on the 2-DE gels only af
ter incubation of the particles in serum. Employing N-terminal microsequenc
ing these protein spots were identified to be fragments of the complement p
rotein C3. Enrichment of these particular spots was most likely a result of
complement activation by the particles. Mechanism of C3 binding to the par
ticle surface and subsequent inactivation by cleavage are discussed in orde
r to explain the results. It could be demonstrated that 2-DE analysis provi
des the possibility to distinguish between adsorption and covalent attachme
nt of C3 to particulate surfaces. The findings indicate that complement act
ivation was caused by covalent binding of the C3 component C3b to the parti
cles' surface. The influence of the incubation medium on the in vitro prote
in adsorption of particulate drug carriers has to be considered when a corr
elation between the protein adsorption pattern and the in vivo behaviour of
the particles is approached. (C) 1999 Elsevier Science Ltd. All rights res
erved.