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Screening for weak monoclonal antibodies in hybridoma technology

Leickt, L LU ; Grubb, A LU orcid and Ohlson, Sten (1998) In Journal of Molecular Recognition 11. p.114-116
Abstract

As the interest in weak-affinity antibodies has been widened by their introduction to various analytical techniques such as HPLC, capillary electrophoresis and biosensors, there has been a need for new screening/monitoring methods. In this study, weak-affinity chromatography was adopted to screen/monitor directly for monoclonal antibodies in ascites. Monoclonal antibodies against a carbohydrate antigen (maltohexaose) were used to evaluate this approach. In short, malthohexaose was immobilized on an HPLC support in such a configuration to allow, during HPLC, retardation of weak monoclonal antibodies. Based on the retention, the affinity or the avidity, as determined by the presence of multiple binding of the monoclonal antibody towards... (More)

As the interest in weak-affinity antibodies has been widened by their introduction to various analytical techniques such as HPLC, capillary electrophoresis and biosensors, there has been a need for new screening/monitoring methods. In this study, weak-affinity chromatography was adopted to screen/monitor directly for monoclonal antibodies in ascites. Monoclonal antibodies against a carbohydrate antigen (maltohexaose) were used to evaluate this approach. In short, malthohexaose was immobilized on an HPLC support in such a configuration to allow, during HPLC, retardation of weak monoclonal antibodies. Based on the retention, the affinity or the avidity, as determined by the presence of multiple binding of the monoclonal antibody towards antigen, can be estimated. In this way it is possible to select clones of hybridomas that produce desired weak monoclonal antibodies. Adjustments in temperature (10-20 degrees C) were used to moderate the retention and hence affinity of the weak monoclonal antibodies during chromatography.

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Please use this url to cite or link to this publication:
author
; and
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Animals, Antibodies, Monoclonal/isolation & purification, Antibody Affinity, Ascites/immunology, Carbohydrate Sequence, Chromatography, Affinity, Chromatography, High Pressure Liquid, Hybridomas/immunology, Mice, Molecular Sequence Data, Oligosaccharides/chemistry
host publication
Journal of Molecular Recognition : Special Issue: Proceedings of the 12th International Symposium on Affinity Interactions - Special Issue: Proceedings of the 12th International Symposium on Affinity Interactions
series title
Journal of Molecular Recognition
volume
11
edition
1-6
pages
114 - 116
external identifiers
  • scopus:0032448546
  • pmid:10076820
ISSN
0952-3499
language
English
LU publication?
yes
id
8f20a070-fc7e-4923-a6ea-3ae465bd6812
alternative location
https://onlinelibrary.wiley.com/doi/abs/10.1002/(SICI)1099-1352(199812)11:1/6%3C114::AID-JMR403%3E3.0.CO;2-%23
date added to LUP
2021-10-29 14:18:26
date last changed
2024-01-12 02:58:08
@inproceedings{8f20a070-fc7e-4923-a6ea-3ae465bd6812,
  abstract     = {{<p>As the interest in weak-affinity antibodies has been widened by their introduction to various analytical techniques such as HPLC, capillary electrophoresis and biosensors, there has been a need for new screening/monitoring methods. In this study, weak-affinity chromatography was adopted to screen/monitor directly for monoclonal antibodies in ascites. Monoclonal antibodies against a carbohydrate antigen (maltohexaose) were used to evaluate this approach. In short, malthohexaose was immobilized on an HPLC support in such a configuration to allow, during HPLC, retardation of weak monoclonal antibodies. Based on the retention, the affinity or the avidity, as determined by the presence of multiple binding of the monoclonal antibody towards antigen, can be estimated. In this way it is possible to select clones of hybridomas that produce desired weak monoclonal antibodies. Adjustments in temperature (10-20 degrees C) were used to moderate the retention and hence affinity of the weak monoclonal antibodies during chromatography.</p>}},
  author       = {{Leickt, L and Grubb, A and Ohlson, Sten}},
  booktitle    = {{Journal of Molecular Recognition : Special Issue: Proceedings of the 12th International Symposium on Affinity Interactions}},
  issn         = {{0952-3499}},
  keywords     = {{Animals; Antibodies, Monoclonal/isolation & purification; Antibody Affinity; Ascites/immunology; Carbohydrate Sequence; Chromatography, Affinity; Chromatography, High Pressure Liquid; Hybridomas/immunology; Mice; Molecular Sequence Data; Oligosaccharides/chemistry}},
  language     = {{eng}},
  pages        = {{114--116}},
  series       = {{Journal of Molecular Recognition}},
  title        = {{Screening for weak monoclonal antibodies in hybridoma technology}},
  url          = {{https://onlinelibrary.wiley.com/doi/abs/10.1002/(SICI)1099-1352(199812)11:1/6%3C114::AID-JMR403%3E3.0.CO;2-%23}},
  volume       = {{11}},
  year         = {{1998}},
}