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Enhanced ELISA sensitivity using TCA for efficient coating of biologically active lipopolysaccharides or lipid A to the solid phase

Hardy, Eugenio; Ohlin, Mats LU and Llano, Manuel (1994) In Journal of Immunological Methods 176(1). p.111-116
Abstract

A new simple, reproducible and sensitive ELISA that uses trichloroacetic acid (TCA) for the coating of lipooligosaccharide (LOS), smooth lipopolysaccharide (LPS) or lipid A to the solid phase has been developed. The experimental parameters (temperature of coating, time of coating, antigen concentration and TCA concentration) were evaluated by a complete factorial design (24). As a result of the evaluation, two main coating procedures were developed. In one, LOS was shown to coat efficiently in 0.2% TCA, at 37°C, when incubated for only 30 min. In the other procedure, LOS in 0.2% TCA was coated at 37°C for 16 h. The slower procedure proved, as expected, to be even more efficient than the former. The new ELISA was compared to... (More)

A new simple, reproducible and sensitive ELISA that uses trichloroacetic acid (TCA) for the coating of lipooligosaccharide (LOS), smooth lipopolysaccharide (LPS) or lipid A to the solid phase has been developed. The experimental parameters (temperature of coating, time of coating, antigen concentration and TCA concentration) were evaluated by a complete factorial design (24). As a result of the evaluation, two main coating procedures were developed. In one, LOS was shown to coat efficiently in 0.2% TCA, at 37°C, when incubated for only 30 min. In the other procedure, LOS in 0.2% TCA was coated at 37°C for 16 h. The slower procedure proved, as expected, to be even more efficient than the former. The new ELISA was compared to three previously reported ELISAs, and showed the greatest sensitivity, probably, as a consequence of the higher coating efficiency of LOS to plates. The biologic activity of LOS was not modified by the low TCA concentration used, as proven by retention of its biological activity in the induction of procoagulant activity in blood mononuclear cells. We conclude that small amounts of biologically active LOS/LPS or lipid A can be coated on solid surfaces by this approach to achieve a rapid and economical assay procedure. © 1994.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Coating efficiency, ELISA, Lipid A, Lipopolysaccharide, Trichloroacetic acid ([76-03-9])
in
Journal of Immunological Methods
volume
176
issue
1
pages
6 pages
publisher
Elsevier
external identifiers
  • Scopus:0028143672
ISSN
0022-1759
DOI
10.1016/0022-1759(94)90355-7
language
English
LU publication?
yes
id
df7b0e53-b912-47be-9f68-fe2b9466c6bd
date added to LUP
2016-04-19 14:12:59
date last changed
2016-04-22 13:22:50
@misc{df7b0e53-b912-47be-9f68-fe2b9466c6bd,
  abstract     = {<p>A new simple, reproducible and sensitive ELISA that uses trichloroacetic acid (TCA) for the coating of lipooligosaccharide (LOS), smooth lipopolysaccharide (LPS) or lipid A to the solid phase has been developed. The experimental parameters (temperature of coating, time of coating, antigen concentration and TCA concentration) were evaluated by a complete factorial design (2<sup>4</sup>). As a result of the evaluation, two main coating procedures were developed. In one, LOS was shown to coat efficiently in 0.2% TCA, at 37°C, when incubated for only 30 min. In the other procedure, LOS in 0.2% TCA was coated at 37°C for 16 h. The slower procedure proved, as expected, to be even more efficient than the former. The new ELISA was compared to three previously reported ELISAs, and showed the greatest sensitivity, probably, as a consequence of the higher coating efficiency of LOS to plates. The biologic activity of LOS was not modified by the low TCA concentration used, as proven by retention of its biological activity in the induction of procoagulant activity in blood mononuclear cells. We conclude that small amounts of biologically active LOS/LPS or lipid A can be coated on solid surfaces by this approach to achieve a rapid and economical assay procedure. © 1994.</p>},
  author       = {Hardy, Eugenio and Ohlin, Mats and Llano, Manuel},
  issn         = {0022-1759},
  keyword      = {Coating efficiency,ELISA,Lipid A,Lipopolysaccharide,Trichloroacetic acid ([76-03-9])},
  language     = {eng},
  month        = {11},
  number       = {1},
  pages        = {111--116},
  publisher    = {ARRAY(0xa425798)},
  series       = {Journal of Immunological Methods},
  title        = {Enhanced ELISA sensitivity using TCA for efficient coating of biologically active lipopolysaccharides or lipid A to the solid phase},
  url          = {http://dx.doi.org/10.1016/0022-1759(94)90355-7},
  volume       = {176},
  year         = {1994},
}