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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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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
Elsevier
external identifiers
  • wos:000242834800019
  • scopus:33845523887
  • pmid:17059411
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
2016-04-01 11:38:45
date last changed
2025-01-14 12:59:00
@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}},
  keywords     = {{latent antithrombin; immunochemical sandwich assay; antithrombin; monoclonal antibody; native-latent antithrombin dimer}},
  language     = {{eng}},
  pages        = {{127--132}},
  publisher    = {{Elsevier}},
  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}},
  doi          = {{10.1111/j.1538-7836.2006.02274.x}},
  volume       = {{5}},
  year         = {{2007}},
}