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New approach to steroid separation based on a low affinity IgM antibody

Strandh, Magnus LU ; Ohlin, Mats LU ; Borrebaeck, Carl A K LU and Ohlson, Sten (1998) In Journal of Immunological Methods 214(1-2). p.73-79
Abstract

IgM antibodies are often of low affinity (dissociation constant (K(d)) > 10-5 M) and therefore they are usually neglected as tools in, e.g., immunoassays. Previous studies have shown that low affinity biological interactions can be studied and exploited in affinity chromatography, biosensor technology and capillary electrophoresis. In this study we have demonstrated that IgM can be a useful ligand for analytical separation of antigens in weak affinity chromatography (WAC). A low affinity human monoclonal IgM antibody, directed at digoxin, was produced in a hybridoma cell culture, purified to homogeneity and immobilized onto an HPLC support. The IgM HPLC column displayed specific weak affinity retention in the 0.01- 0.1 mM... (More)

IgM antibodies are often of low affinity (dissociation constant (K(d)) > 10-5 M) and therefore they are usually neglected as tools in, e.g., immunoassays. Previous studies have shown that low affinity biological interactions can be studied and exploited in affinity chromatography, biosensor technology and capillary electrophoresis. In this study we have demonstrated that IgM can be a useful ligand for analytical separation of antigens in weak affinity chromatography (WAC). A low affinity human monoclonal IgM antibody, directed at digoxin, was produced in a hybridoma cell culture, purified to homogeneity and immobilized onto an HPLC support. The IgM HPLC column displayed specific weak affinity retention in the 0.01- 0.1 mM range as evaluated with digoxin and ouabain. The specificity was not affected when samples of ouabain in a crude environment of diluted serum were separated on the IgM column. These findings suggest an approach in immunoadsorbent technology where biomolecules can be analyzed and separated with weak affinity chromatography using IgM as a general affinity ligand.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Digoxin, Low affinity monoclonal IgM antibody, Weak affinity chromatography (WAC)
in
Journal of Immunological Methods
volume
214
issue
1-2
pages
7 pages
publisher
Elsevier
external identifiers
  • Scopus:0031813293
ISSN
0022-1759
DOI
10.1016/S0022-1759(98)00039-8
language
English
LU publication?
yes
id
eac32f4e-e443-478b-991c-49e02a2ae3fd
date added to LUP
2016-04-19 14:06:06
date last changed
2016-10-13 05:06:40
@misc{eac32f4e-e443-478b-991c-49e02a2ae3fd,
  abstract     = {<p>IgM antibodies are often of low affinity (dissociation constant (K(d)) &gt; 10<sup>-5</sup> M) and therefore they are usually neglected as tools in, e.g., immunoassays. Previous studies have shown that low affinity biological interactions can be studied and exploited in affinity chromatography, biosensor technology and capillary electrophoresis. In this study we have demonstrated that IgM can be a useful ligand for analytical separation of antigens in weak affinity chromatography (WAC). A low affinity human monoclonal IgM antibody, directed at digoxin, was produced in a hybridoma cell culture, purified to homogeneity and immobilized onto an HPLC support. The IgM HPLC column displayed specific weak affinity retention in the 0.01- 0.1 mM range as evaluated with digoxin and ouabain. The specificity was not affected when samples of ouabain in a crude environment of diluted serum were separated on the IgM column. These findings suggest an approach in immunoadsorbent technology where biomolecules can be analyzed and separated with weak affinity chromatography using IgM as a general affinity ligand.</p>},
  author       = {Strandh, Magnus and Ohlin, Mats and Borrebaeck, Carl A K and Ohlson, Sten},
  issn         = {0022-1759},
  keyword      = {Digoxin,Low affinity monoclonal IgM antibody,Weak affinity chromatography (WAC)},
  language     = {eng},
  month        = {05},
  number       = {1-2},
  pages        = {73--79},
  publisher    = {ARRAY(0xafa4420)},
  series       = {Journal of Immunological Methods},
  title        = {New approach to steroid separation based on a low affinity IgM antibody},
  url          = {http://dx.doi.org/10.1016/S0022-1759(98)00039-8},
  volume       = {214},
  year         = {1998},
}