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Spot immunoprecipitate assay in gel compared with immune reactions in solution and applied to antibody titration

Wadsworth, C; Nilsson, L A and Bjursten, Lars Magnus LU (1983) In Journal of Immunological Methods 62(2). p.217-229
Abstract
Precipitation profiles of spot immunoprecipitate assay (SIA) reactions in gel are shown to have characteristics analogous to classic precipitin curves representing such immune reactions in solution. Thus SIA profiles of precipitated human albumin (HSA) and anti-HSA as checkerboard analysis permit: (1) determination of optimal proportions of antibody and antigen, (2) titration of antiserum as to antigen-binding capacity and (3) estimation of the concentration of specific antibody in an antiserum. The validity of the method was confirmed by similar results already reported for precipitin analysis of anti-HSA in solution. The stained protein assay method for determination of total protein used for the calculations had a reliable (r = 0.9979)... (More)
Precipitation profiles of spot immunoprecipitate assay (SIA) reactions in gel are shown to have characteristics analogous to classic precipitin curves representing such immune reactions in solution. Thus SIA profiles of precipitated human albumin (HSA) and anti-HSA as checkerboard analysis permit: (1) determination of optimal proportions of antibody and antigen, (2) titration of antiserum as to antigen-binding capacity and (3) estimation of the concentration of specific antibody in an antiserum. The validity of the method was confirmed by similar results already reported for precipitin analysis of anti-HSA in solution. The stained protein assay method for determination of total protein used for the calculations had a reliable (r = 0.9979) working range for quantification between 0.1 and 0.83 microgram protein in 3 microliters samples (greater than or equal to 0.03 g/l). Study of SIA profiles in the extreme antibody excess region confirmed the validity of SIA quantification. The high sensitivity of SIA as compared with radial immunodiffusion is related to high ratios (8-9) of antibody to antigen; the reliable detection limit for SIA is 10 mg/l in a 3 microliters sample. SIA is easy to perform with simple laboratory equipment, allows measurement of any precipitable antigen with very small amounts of reactants, and gives results within an hour. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
antibody titration, precipitation in gel, optimal proportion of antigen and antibody, antigen-antibody reaction
in
Journal of Immunological Methods
volume
62
issue
2
pages
217 - 229
publisher
Elsevier
external identifiers
  • pmid:6886433
  • scopus:0020511157
ISSN
1872-7905
DOI
10.1016/0022-1759(83)90249-1
language
English
LU publication?
yes
id
89167044-848f-49b2-8efb-a58a116a8d2d (old id 1103001)
date added to LUP
2008-08-13 11:09:39
date last changed
2017-01-01 06:40:44
@article{89167044-848f-49b2-8efb-a58a116a8d2d,
  abstract     = {Precipitation profiles of spot immunoprecipitate assay (SIA) reactions in gel are shown to have characteristics analogous to classic precipitin curves representing such immune reactions in solution. Thus SIA profiles of precipitated human albumin (HSA) and anti-HSA as checkerboard analysis permit: (1) determination of optimal proportions of antibody and antigen, (2) titration of antiserum as to antigen-binding capacity and (3) estimation of the concentration of specific antibody in an antiserum. The validity of the method was confirmed by similar results already reported for precipitin analysis of anti-HSA in solution. The stained protein assay method for determination of total protein used for the calculations had a reliable (r = 0.9979) working range for quantification between 0.1 and 0.83 microgram protein in 3 microliters samples (greater than or equal to 0.03 g/l). Study of SIA profiles in the extreme antibody excess region confirmed the validity of SIA quantification. The high sensitivity of SIA as compared with radial immunodiffusion is related to high ratios (8-9) of antibody to antigen; the reliable detection limit for SIA is 10 mg/l in a 3 microliters sample. SIA is easy to perform with simple laboratory equipment, allows measurement of any precipitable antigen with very small amounts of reactants, and gives results within an hour.},
  author       = {Wadsworth, C and Nilsson, L A and Bjursten, Lars Magnus},
  issn         = {1872-7905},
  keyword      = {antibody titration,precipitation in gel,optimal proportion of antigen and antibody,antigen-antibody reaction},
  language     = {eng},
  number       = {2},
  pages        = {217--229},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Spot immunoprecipitate assay in gel compared with immune reactions in solution and applied to antibody titration},
  url          = {http://dx.doi.org/10.1016/0022-1759(83)90249-1},
  volume       = {62},
  year         = {1983},
}