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A hemolytic assay for the estimation of functional mannose-binding lectin levels in human serum

Kuipers, S; Aerts, PC; Sjöholm, Anders LU ; Harmsen, T and van Dijk, H (2002) In Journal of Immunological Methods 268(2). p.149-157
Abstract
A simple assay was developed to estimate functional mannose-binding lectin (MBL) levels in serum based on the principle of yeast-induced bystander lysis of chicken erythrocytes (ChE). The assay is sensitive to inhibition by ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) (which allows alternative pathway activation), ethylene diamine tetraacetic acid (EDTA), mannose, N-acetylglucosamine and C1 esterase inhibitor (C1-INH), whereas it was not inhibited by galactose. A high-titer human anti-mannan antibody-containing serum with 0.06 mug MBL/ml gave a functional signal corresponding to 0.12 mug equivalents MBL/ml, indicating that anti-mannan antibodies are poorly hemolytic in the assay. The assay is well suited... (More)
A simple assay was developed to estimate functional mannose-binding lectin (MBL) levels in serum based on the principle of yeast-induced bystander lysis of chicken erythrocytes (ChE). The assay is sensitive to inhibition by ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) (which allows alternative pathway activation), ethylene diamine tetraacetic acid (EDTA), mannose, N-acetylglucosamine and C1 esterase inhibitor (C1-INH), whereas it was not inhibited by galactose. A high-titer human anti-mannan antibody-containing serum with 0.06 mug MBL/ml gave a functional signal corresponding to 0.12 mug equivalents MBL/ml, indicating that anti-mannan antibodies are poorly hemolytic in the assay. The assay is well suited for the large-scale testing of patient samples for a functional MBL pathway of complement activation. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
bystander hemolysis, functional assay, MBL, lectin pathway, Saccharomyces cerevisiae
in
Journal of Immunological Methods
volume
268
issue
2
pages
149 - 157
publisher
Elsevier
external identifiers
  • pmid:12215383
  • wos:000178241400004
  • scopus:0037107536
ISSN
1872-7905
DOI
10.1016/S0022-1759(02)00192-8
language
English
LU publication?
yes
id
9b9f08dc-e443-4c3f-bbd4-147768c35087 (old id 328321)
date added to LUP
2007-11-05 13:19:37
date last changed
2017-01-01 06:46:24
@article{9b9f08dc-e443-4c3f-bbd4-147768c35087,
  abstract     = {A simple assay was developed to estimate functional mannose-binding lectin (MBL) levels in serum based on the principle of yeast-induced bystander lysis of chicken erythrocytes (ChE). The assay is sensitive to inhibition by ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) (which allows alternative pathway activation), ethylene diamine tetraacetic acid (EDTA), mannose, N-acetylglucosamine and C1 esterase inhibitor (C1-INH), whereas it was not inhibited by galactose. A high-titer human anti-mannan antibody-containing serum with 0.06 mug MBL/ml gave a functional signal corresponding to 0.12 mug equivalents MBL/ml, indicating that anti-mannan antibodies are poorly hemolytic in the assay. The assay is well suited for the large-scale testing of patient samples for a functional MBL pathway of complement activation.},
  author       = {Kuipers, S and Aerts, PC and Sjöholm, Anders and Harmsen, T and van Dijk, H},
  issn         = {1872-7905},
  keyword      = {bystander hemolysis,functional assay,MBL,lectin pathway,Saccharomyces cerevisiae},
  language     = {eng},
  number       = {2},
  pages        = {149--157},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {A hemolytic assay for the estimation of functional mannose-binding lectin levels in human serum},
  url          = {http://dx.doi.org/10.1016/S0022-1759(02)00192-8},
  volume       = {268},
  year         = {2002},
}