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A novel method for direct measurement of complement convertases activity in human serum.

Blom, Anna LU ; Volokhina, Elena B; Fransson, Viktor LU ; Strömberg, Patrik; Berghard, Lotta; Viktorelius, Margareta; Mollnes, Tom Eirik; Lopez-Holmberg, Margarita; van den Heuvel, Lambertus P and Goodship, Tim H, et al. (2014) In Clinical and Experimental Immunology 178(1). p.142-153
Abstract
Complement convertases are enzymatic complexes that play a central role in sustaining and amplification of the complement cascade. Impairment of complement function directly or indirectly leads to pathologic conditions including higher infection rate, kidney diseases, autoimmune- or neurodegenerative diseases and ischemia-reperfusion injury. An assay for direct measurement of activity of the convertases in patient sera is not available. Existing assays testing convertase function are based on purified complement components and thus, convertase formation occurs under non-physiological conditions. We designed a new assay, in which C5 blocking compounds enabled separation of the complement cascade into two phases: the first ending at the... (More)
Complement convertases are enzymatic complexes that play a central role in sustaining and amplification of the complement cascade. Impairment of complement function directly or indirectly leads to pathologic conditions including higher infection rate, kidney diseases, autoimmune- or neurodegenerative diseases and ischemia-reperfusion injury. An assay for direct measurement of activity of the convertases in patient sera is not available. Existing assays testing convertase function are based on purified complement components and thus, convertase formation occurs under non-physiological conditions. We designed a new assay, in which C5 blocking compounds enabled separation of the complement cascade into two phases: the first ending at the stage of C5 convertases and the second ending with membrane attack complex formation. Use of rabbit erythrocytes or antibody-sensitized sheep erythrocytes as the platforms for convertase formation enabled easy readout based on measurement of hemolysis. Thus, properties of patient sera could be studied directly regarding convertase activity and membrane attack complex formation. Another advantage of this assay was the possibility to screen for host factors such as C3 nephritic factor and other anti-complement autoantibodies, or gain-of-function mutations, which prolong half-live of complement convertases. Herein, we present proof of concept, detailed description and validation of this novel assay. (Less)
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Contribution to journal
publication status
published
subject
in
Clinical and Experimental Immunology
volume
178
issue
1
pages
142 - 153
publisher
British Society for Immunology
external identifiers
  • pmid:24853370
  • wos:000342914100017
  • scopus:84908212932
ISSN
0009-9104
DOI
10.1111/cei.12388
language
English
LU publication?
yes
id
37e299b8-954e-4eb3-bf61-a7826947c937 (old id 4452711)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/24853370?dopt=Abstract
date added to LUP
2014-06-05 00:14:32
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2017-03-26 03:11:38
@article{37e299b8-954e-4eb3-bf61-a7826947c937,
  abstract     = {Complement convertases are enzymatic complexes that play a central role in sustaining and amplification of the complement cascade. Impairment of complement function directly or indirectly leads to pathologic conditions including higher infection rate, kidney diseases, autoimmune- or neurodegenerative diseases and ischemia-reperfusion injury. An assay for direct measurement of activity of the convertases in patient sera is not available. Existing assays testing convertase function are based on purified complement components and thus, convertase formation occurs under non-physiological conditions. We designed a new assay, in which C5 blocking compounds enabled separation of the complement cascade into two phases: the first ending at the stage of C5 convertases and the second ending with membrane attack complex formation. Use of rabbit erythrocytes or antibody-sensitized sheep erythrocytes as the platforms for convertase formation enabled easy readout based on measurement of hemolysis. Thus, properties of patient sera could be studied directly regarding convertase activity and membrane attack complex formation. Another advantage of this assay was the possibility to screen for host factors such as C3 nephritic factor and other anti-complement autoantibodies, or gain-of-function mutations, which prolong half-live of complement convertases. Herein, we present proof of concept, detailed description and validation of this novel assay.},
  author       = {Blom, Anna and Volokhina, Elena B and Fransson, Viktor and Strömberg, Patrik and Berghard, Lotta and Viktorelius, Margareta and Mollnes, Tom Eirik and Lopez-Holmberg, Margarita and van den Heuvel, Lambertus P and Goodship, Tim H and Marchbank, Kevin J and Okroj, Marcin},
  issn         = {0009-9104},
  language     = {eng},
  number       = {1},
  pages        = {142--153},
  publisher    = {British Society for Immunology},
  series       = {Clinical and Experimental Immunology},
  title        = {A novel method for direct measurement of complement convertases activity in human serum.},
  url          = {http://dx.doi.org/10.1111/cei.12388},
  volume       = {178},
  year         = {2014},
}