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Affinity screening for weak monoclonal antibodies

Leickt, Lisa LU ; Grubb, Anders LU orcid and Ohlson, Sten (1998) In Journal of Immunological Methods 220(1-2). p.19-24
Abstract

When selecting for monoclonal antibodies of a desired affinity, affinity chromatography can be a feasible alternative. This is of particular interest when low affinity monoclonal antibodies (dissociation constant (Kd) > 10(-4) M) are screened, as they are not easily recognised by traditional immunoassay procedures. In this study we have evaluated this approach by monitoring low affinity monoclonal antibodies on high performance liquid affinity chromatography columns with oligosaccharides, dinitrophenol and digoxin as immobilised antigen. Crude monoclonal antibodies in ascites or cell culture supematants, directed against these antigens, were retarded or adsorbed according to affinity or avidity on the antigen columns. Based on... (More)

When selecting for monoclonal antibodies of a desired affinity, affinity chromatography can be a feasible alternative. This is of particular interest when low affinity monoclonal antibodies (dissociation constant (Kd) > 10(-4) M) are screened, as they are not easily recognised by traditional immunoassay procedures. In this study we have evaluated this approach by monitoring low affinity monoclonal antibodies on high performance liquid affinity chromatography columns with oligosaccharides, dinitrophenol and digoxin as immobilised antigen. Crude monoclonal antibodies in ascites or cell culture supematants, directed against these antigens, were retarded or adsorbed according to affinity or avidity on the antigen columns. Based on antibody retention, we were able to select hybridomas with the desired low affinity characteristics.

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keywords
Animals, Antibodies, Monoclonal/immunology, Antibody Affinity, Ascitic Fluid/immunology, Chromatography, Affinity, Chromatography, High Pressure Liquid, Digoxin/immunology, Dinitrophenols/immunology, Hemocyanins/immunology, Immunoglobulin M/immunology, Mice, Oligosaccharides/immunology
in
Journal of Immunological Methods
volume
220
issue
1-2
pages
19 - 24
publisher
Elsevier
external identifiers
  • scopus:0031733261
  • pmid:9839922
ISSN
0022-1759
DOI
10.1016/s0022-1759(98)00163-x
language
English
LU publication?
yes
id
277e7735-c310-480e-be51-ae128edd5847
date added to LUP
2021-11-02 07:58:36
date last changed
2024-01-12 02:57:25
@article{277e7735-c310-480e-be51-ae128edd5847,
  abstract     = {{<p>When selecting for monoclonal antibodies of a desired affinity, affinity chromatography can be a feasible alternative. This is of particular interest when low affinity monoclonal antibodies (dissociation constant (Kd) &gt; 10(-4) M) are screened, as they are not easily recognised by traditional immunoassay procedures. In this study we have evaluated this approach by monitoring low affinity monoclonal antibodies on high performance liquid affinity chromatography columns with oligosaccharides, dinitrophenol and digoxin as immobilised antigen. Crude monoclonal antibodies in ascites or cell culture supematants, directed against these antigens, were retarded or adsorbed according to affinity or avidity on the antigen columns. Based on antibody retention, we were able to select hybridomas with the desired low affinity characteristics.</p>}},
  author       = {{Leickt, Lisa and Grubb, Anders and Ohlson, Sten}},
  issn         = {{0022-1759}},
  keywords     = {{Animals; Antibodies, Monoclonal/immunology; Antibody Affinity; Ascitic Fluid/immunology; Chromatography, Affinity; Chromatography, High Pressure Liquid; Digoxin/immunology; Dinitrophenols/immunology; Hemocyanins/immunology; Immunoglobulin M/immunology; Mice; Oligosaccharides/immunology}},
  language     = {{eng}},
  number       = {{1-2}},
  pages        = {{19--24}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Immunological Methods}},
  title        = {{Affinity screening for weak monoclonal antibodies}},
  url          = {{http://dx.doi.org/10.1016/s0022-1759(98)00163-x}},
  doi          = {{10.1016/s0022-1759(98)00163-x}},
  volume       = {{220}},
  year         = {{1998}},
}