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An immunochemical method for quantitative determination of latent antithrombin, the reactive center loop-inserted uncleaved form of antithrombin.

Kjellberg, Margareta LU ; Rimac, Berit LU and Stenflo, Johan LU (2007) In Journal of Thrombosis and Haemostasis 5. p.127-132
Abstract
Antithrombin (AT) is a serine protease inhibitor that has thrombin, factors IXa and Xa as target proteases. In addition to active native AT, two other forms have been identified in plasma: the reactive center loop inserted cleaved and latent, uncleaved forms. Both have been shown to be present in normal human blood. Latent AT forms a dimer with native AT in vitro, thus inactivating the native form. Here we describe a mouse monoclonal antibody, 8C8, that is specific for latent AT. The affinity of 8C8 was found to be 500-fold higher for latent than for native AT and 5000-fold higher for latent than for cleaved AT. A sandwich assay was developed to measure the concentration of latent AT in plasma, which was found to be similar to 4.8 mg L-1... (More)
Antithrombin (AT) is a serine protease inhibitor that has thrombin, factors IXa and Xa as target proteases. In addition to active native AT, two other forms have been identified in plasma: the reactive center loop inserted cleaved and latent, uncleaved forms. Both have been shown to be present in normal human blood. Latent AT forms a dimer with native AT in vitro, thus inactivating the native form. Here we describe a mouse monoclonal antibody, 8C8, that is specific for latent AT. The affinity of 8C8 was found to be 500-fold higher for latent than for native AT and 5000-fold higher for latent than for cleaved AT. A sandwich assay was developed to measure the concentration of latent AT in plasma, which was found to be similar to 4.8 mg L-1 in healthy individuals. The K-D of the interaction between native and latent AT was found to be 51 mu M, i.e. far above the plasma concentration of both native and latent AT, indicating a negligible complex formation in blood. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
latent antithrombin, immunochemical sandwich assay, antithrombin, monoclonal antibody, native-latent antithrombin dimer
in
Journal of Thrombosis and Haemostasis
volume
5
pages
127 - 132
publisher
Federation of European Neuroscience Societies and Blackwell Publishing Ltd
external identifiers
  • wos:000242834800019
  • scopus:33845523887
ISSN
1538-7933
DOI
10.1111/j.1538-7836.2006.02274.x
language
English
LU publication?
yes
id
b3d04725-411c-4cef-b1e9-7f5330364e77 (old id 162189)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17059411&dopt=Abstract
date added to LUP
2007-07-25 15:14:39
date last changed
2017-01-01 04:24:10
@article{b3d04725-411c-4cef-b1e9-7f5330364e77,
  abstract     = {Antithrombin (AT) is a serine protease inhibitor that has thrombin, factors IXa and Xa as target proteases. In addition to active native AT, two other forms have been identified in plasma: the reactive center loop inserted cleaved and latent, uncleaved forms. Both have been shown to be present in normal human blood. Latent AT forms a dimer with native AT in vitro, thus inactivating the native form. Here we describe a mouse monoclonal antibody, 8C8, that is specific for latent AT. The affinity of 8C8 was found to be 500-fold higher for latent than for native AT and 5000-fold higher for latent than for cleaved AT. A sandwich assay was developed to measure the concentration of latent AT in plasma, which was found to be similar to 4.8 mg L-1 in healthy individuals. The K-D of the interaction between native and latent AT was found to be 51 mu M, i.e. far above the plasma concentration of both native and latent AT, indicating a negligible complex formation in blood.},
  author       = {Kjellberg, Margareta and Rimac, Berit and Stenflo, Johan},
  issn         = {1538-7933},
  keyword      = {latent antithrombin,immunochemical sandwich assay,antithrombin,monoclonal antibody,native-latent antithrombin dimer},
  language     = {eng},
  pages        = {127--132},
  publisher    = {Federation of European Neuroscience Societies and Blackwell Publishing Ltd},
  series       = {Journal of Thrombosis and Haemostasis},
  title        = {An immunochemical method for quantitative determination of latent antithrombin, the reactive center loop-inserted uncleaved form of antithrombin.},
  url          = {http://dx.doi.org/10.1111/j.1538-7836.2006.02274.x},
  volume       = {5},
  year         = {2007},
}