Effect of time, temperature and anticoagulants on in vitro complement activation: consequences for collection and preservation of samples to be examined for complement activation.
The effects of time, temperature, ethylene-diamine-tetra-acetic acid (EDTA), citrate and heparin on in vitro complement activation were examined in enzyme immuno assays (EIA) for detection of C3 activation products and the terminal complement complex (TCC). In vitro complement activation occurred during coagulation since baseline concentrations of activation products were considerably higher in serum than in plasma. EDTA was more efficient than citrate and heparin in inhibiting in vitro activation. Minimal activation was observed in all preparations when samples were kept at 4 degrees C for up to ten days, whereas a very rapid increase in activation products occurred even in EDTA plasma when the temperature was elevated. Based on the data obtained, guidelines for the collection and preservation of samples to be examined for complement activation are given.
Mollnes
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TE
Institute of Immunology and Rheumatology, National Hospital, Oslo, Norway.
Garred
P
P
Bergseth
G
G
D001793:Blood Preservation; D002951:Citrates / Q000494:pharmacology; D019343:Citric Acid; D003167:Complement Activation / Q000187:drug effects; D003176:Complement C3 / Q000378:metabolism; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000096:biosynthesis; D004492:Edetic Acid / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005615:Freezing; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature
D002951:Citrates; D003176:Complement C3; D015938:Complement Membrane Attack Complex; D000939:Epitopes; D019343:Citric Acid; D006493:Heparin; D003165:Complement System Proteins; D004492:Edetic Acid