Ellipsometric in vitro studies on the activation of complement by human immunoglobulins M and G after adsorption to methylated silicon

• Human serum immunoglobulin M (IgM) or human immunoglobulin G (IgG) were adsorbed to dichlorodimethyl silane (DDS) treated silicon. Subsequently, the model surfaces were incubated in normal-, complement factor 1q (C1q)-complement factor B or complement factor 2 (C2)-depleted human sera at 37°C for up to 1.5 h. The serum deposition and binding of selected polyclonal complement antibodies into this layer were then quantified by null ellipsometry. Both types of precoated surfaces bound large amounts of anti-complement factor 3c (anti-C3c), anti-properdin and anti-C3d, after incubation in normal serum. In contrast to IgG coated surfaces, IgM coated surfaces bound no anti-C1q after the serum incubations and no anti-C3c deposition lag time was observed after incubations in EGTA serum. Upon immersions of IgM coated surfaces in the different sera, a rapid complement activation via a C1q factor B, and Ca2+-independent, but C2 dependent pathway, was indicated. When IgM was instead immobilized to APTES/glutaraldehyde surfaces, anti-C3c deposition was lower after incubations in EGTA than normal serum. The results suggest that, under the present experimental conditions, human IgM and IgG activate the complement system differently. (C) 2001 Elsevier Science B.V.

Keyword

Activation

Complement

Ellipsometry

Immunoglobulin M

Kinetics

TECHNOLOGY

TEKNIKVETENSKAP

Publication and Content Type

ref (subject category)

art (subject category)