Isolation and characterization of an inter-alpha-trypsin inhibitor-IgG complex from human serum.
Inter-alpha-trypsin inhibitor is a human serum protease inhibitor of Mr 180 000 which may release physiological derivatives. A complex between IgG and an inter-alpha-trypsin inhibitor derivative of Mr 30 000 has been recently detected in human serum and was found to be inactive against trypsin, in contrast with the known inhibitory activity of the free 30-kDa derivative. The present study deals with detailed characterization of an inter-alpha-trypsin inhibitor-IgG complex following its purification by affinity chromatography techniques (anti-inter-alpha-trypsin inhibitor immunoadsorbent and Protein A-Sepharose) in mild conditions. The resulting product reacted simultaneously with anti-IgG and anti-inter-alpha-trypsin inhibitor antibodies. This complex contained Mr 180 000 inhibitor at least to some extent. It migrated in the beta-gamma zone in agarose; its molecular weight was estimated to be 1 500 000 or more; part of it displayed covalent bonding between inter-alpha-trypsin inhibitor and IgG; it had a trypsin inhibitor activity. Immunoelectrophoresis allowed one to demonstrate the native complex in serum owing to the use of anti-inter-alpha-trypsin inhibitor and anti-gamma radioactively labelled antibodies. The double immunoreactivity thus evidenced proved to be heterogeneous with respect to its level and location in the native as well as in the purified complex.
D000510:Alpha-Globulins / Q000302:isolation & purification*; D006801:Humans; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008970:Molecular Weight; D011485:Protein Binding; D014361:Trypsin Inhibitors / Q000302:isolation & purification*
D000510:Alpha-Globulins; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D014361:Trypsin Inhibitors; C011877:inter-alpha-inhibitor