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Flow cytometric analysis of the stability of antibody production by human × human × mouse heterohybridoma subclones

Ohlin, Mats LU orcid and Borrebaeck, Carl A K LU (1994) In Journal of Immunological Methods 170(1). p.75-82
Abstract

Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human × human × mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of heavy chain was found to correlate with low antibody productivity. However, the majority of cells were not completely devoid of heavy chains suggesting that the genetic information for the γ chain was not lost. In contrast, the gene encoding the κ light chain was shown to be absent from the subclones expressing low levels of heavy chain and these subclones also contained substantially reduced levels of heavy chain mRNA,... (More)

Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human × human × mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of heavy chain was found to correlate with low antibody productivity. However, the majority of cells were not completely devoid of heavy chains suggesting that the genetic information for the γ chain was not lost. In contrast, the gene encoding the κ light chain was shown to be absent from the subclones expressing low levels of heavy chain and these subclones also contained substantially reduced levels of heavy chain mRNA, suggesting that the production of this protein was controlled at the level of transcription or mRNA stability. In conclusion, the correlation of staining intensity as observed by flow cytometry and antibody productivity makes flow cytometry a suitable technique for the rapid evaluation of heterohybridoma cell lines used for antibody production.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
keywords
Flow cytometry, Instability, Monoclonal antibody, human, Productivity
in
Journal of Immunological Methods
volume
170
issue
1
pages
8 pages
publisher
Elsevier
external identifiers
  • pmid:8157990
  • scopus:0028216903
ISSN
0022-1759
DOI
10.1016/0022-1759(94)90247-X
language
English
LU publication?
yes
id
c457fc2a-bf9b-45a6-9d56-9a5a110e8e12
date added to LUP
2016-04-19 14:15:29
date last changed
2024-01-03 23:53:33
@article{c457fc2a-bf9b-45a6-9d56-9a5a110e8e12,
  abstract     = {{<p>Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human × human × mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of heavy chain was found to correlate with low antibody productivity. However, the majority of cells were not completely devoid of heavy chains suggesting that the genetic information for the γ chain was not lost. In contrast, the gene encoding the κ light chain was shown to be absent from the subclones expressing low levels of heavy chain and these subclones also contained substantially reduced levels of heavy chain mRNA, suggesting that the production of this protein was controlled at the level of transcription or mRNA stability. In conclusion, the correlation of staining intensity as observed by flow cytometry and antibody productivity makes flow cytometry a suitable technique for the rapid evaluation of heterohybridoma cell lines used for antibody production.</p>}},
  author       = {{Ohlin, Mats and Borrebaeck, Carl A K}},
  issn         = {{0022-1759}},
  keywords     = {{Flow cytometry; Instability; Monoclonal antibody, human; Productivity}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{1}},
  pages        = {{75--82}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Immunological Methods}},
  title        = {{Flow cytometric analysis of the stability of antibody production by human × human × mouse heterohybridoma subclones}},
  url          = {{http://dx.doi.org/10.1016/0022-1759(94)90247-X}},
  doi          = {{10.1016/0022-1759(94)90247-X}},
  volume       = {{170}},
  year         = {{1994}},
}