Skip to main content
NCBI home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1988 Sep;73(3):484–488.

Effect of time, temperature and anticoagulants on in vitro complement activation: consequences for collection and preservation of samples to be examined for complement activation.

T E Mollnes 1, P Garred 1, G Bergseth 1
PMCID: PMC1541764  PMID: 2463123

Abstract

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.

Full text

PDF
484

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Garred P., Mollnes T. E., Lea T., Fischer E. Characterization of a monoclonal antibody MoAb bH6 reacting with a neoepitope of human C3 expressed on C3b, iC3b, and C3c. Scand J Immunol. 1988 Mar;27(3):319–327. doi: 10.1111/j.1365-3083.1988.tb02353.x. [DOI] [PubMed] [Google Scholar]
  2. Garred P., Mollnes T. E., Lea T. Quantification in enzyme-linked immunosorbent assay of a C3 neoepitope expressed on activated human complement factor C3. Scand J Immunol. 1988 Mar;27(3):329–335. doi: 10.1111/j.1365-3083.1988.tb02354.x. [DOI] [PubMed] [Google Scholar]
  3. Kazatchkine M. D., Fearon D. T., Metcalfe D. D., Rosenberg R. D., Austen K. F. Structural determinants of the capacity of heparin to inhibit the formation of the human amplification C3 convertase. J Clin Invest. 1981 Jan;67(1):223–228. doi: 10.1172/JCI110017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Lachmann P. J., Pangburn M. K., Oldroyd R. G. Breakdown of C3 after complement activation. Identification of a new fragment C3g, using monoclonal antibodies. J Exp Med. 1982 Jul 1;156(1):205–216. doi: 10.1084/jem.156.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Mollnes T. E. Early- and late-phase activation of complement evaluated by plasma levels of C3d,g and the terminal complement complex. Complement. 1985;2(2-3):156–164. doi: 10.1159/000467856. [DOI] [PubMed] [Google Scholar]
  6. Mollnes T. E., Lachmann P. J. Activation of the third component of complement (C3) detected by a monoclonal anti-C3'g' neoantigen antibody in a one-step enzyme immunoassay. J Immunol Methods. 1987 Aug 3;101(2):201–207. doi: 10.1016/0022-1759(87)90151-7. [DOI] [PubMed] [Google Scholar]
  7. Mollnes T. E., Lea T., Frøland S. S., Harboe M. Quantification of the terminal complement complex in human plasma by an enzyme-linked immunosorbent assay based on monoclonal antibodies against a neoantigen of the complex. Scand J Immunol. 1985 Aug;22(2):197–202. doi: 10.1111/j.1365-3083.1985.tb01871.x. [DOI] [PubMed] [Google Scholar]
  8. Mollnes T. E., Ng Y. C., Peters D. K., Lea T., Tschopp J., Harboe M. Effect of nephritic factor on C3 and on the terminal pathway of complement in vivo and in vitro. Clin Exp Immunol. 1986 Jul;65(1):73–79. [PMC free article] [PubMed] [Google Scholar]
  10. Pangburn M. K., Schreiber R. D., Müller-Eberhard H. J. Formation of the initial C3 convertase of the alternative complement pathway. Acquisition of C3b-like activities by spontaneous hydrolysis of the putative thioester in native C3. J Exp Med. 1981 Sep 1;154(3):856–867. doi: 10.1084/jem.154.3.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Samuel D. J., Amlot P. L., Shepherd P., Lachmann P. J. An efficient one-step method for isolating immune complexes from whole serum using a monoclonal anti-C3g affinity immunosorbent. Clin Exp Immunol. 1986 Aug;65(2):458–464. [PMC free article] [PubMed] [Google Scholar]
  12. Weiler J. M., Yurt R. W., Fearon D. T., Austen K. F. Modulation of the formation of the amplification convertase of complement, C3b, Bb, by native and commercial heparin. J Exp Med. 1978 Feb 1;147(2):409–421. doi: 10.1084/jem.147.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ziccardi R. J. Spontaneous activation of the first component of human complement (C1) by an intramolecular autocatalytic mechanism. J Immunol. 1982 Jun;128(6):2500–2504. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES