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Cross-reacting monoclonal anti-alpha 1-microglobulin antibodies produced by multi-species immunization and using protein G for the screening assay

Nilson, B LU ; Akerström, B LU and Lögdberg, L (1987) In Journal of Immunological Methods 99(1). p.39-45
Abstract

In order to generate monoclonal antibodies (MAb) directed against the low molecular weight glycoprotein alpha 1-microglobulin, a BALB/c mouse was immunized with a mixture of human, guinea pig, rat and rabbit alpha 1-microglobulin homologues (multi-species immunization) and boosted several times. On day 194, the mouse splenocytes were fused to SP2/0 myeloma cells. The resulting hybridomas were screened for anti-alpha 1-microglobulin activity against the alpha 1-microglobulin mixture or against the individual homologues. For this screening, protein G (the newly described IgG-binding streptococcal protein) was used in a solid-phase radioimmunoassay. The binding of protein G to immobilized antigen-antibody complexes was enhanced by... (More)

In order to generate monoclonal antibodies (MAb) directed against the low molecular weight glycoprotein alpha 1-microglobulin, a BALB/c mouse was immunized with a mixture of human, guinea pig, rat and rabbit alpha 1-microglobulin homologues (multi-species immunization) and boosted several times. On day 194, the mouse splenocytes were fused to SP2/0 myeloma cells. The resulting hybridomas were screened for anti-alpha 1-microglobulin activity against the alpha 1-microglobulin mixture or against the individual homologues. For this screening, protein G (the newly described IgG-binding streptococcal protein) was used in a solid-phase radioimmunoassay. The binding of protein G to immobilized antigen-antibody complexes was enhanced by pre-incubation with rabbit anti-mouse immunoglobulin G. The result was a panel of nine established hybridoma lines, all producing unique monoclonal antibodies, of IgG1 or IgG2a class, to alpha 1-microglobulin. The antibodies were not only reactive in solid-phase radioimmunoassay, but they could also immunoprecipitate 125I-labeled soluble alpha 1-microglobulin. Moreover, they reacted specifically with the alpha 1-microglobulin band in Western blots of urinary proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Such monoclonal antibodies are potentially valuable reagents for the further characterization of alpha 1-microglobulin.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Alpha-Globulins/immunology, Animals, Antibodies, Monoclonal/biosynthesis, Cross Reactions, Guinea Pigs, Humans, Hybridomas/analysis, Immunization/methods, Mice, Mice, Inbred BALB C, Nerve Tissue Proteins, Radioimmunoassay/methods, Rats
in
Journal of Immunological Methods
volume
99
issue
1
pages
39 - 45
publisher
Elsevier
external identifiers
  • pmid:2437206
  • scopus:0023233859
ISSN
0022-1759
DOI
10.1016/0022-1759(87)90029-9
language
English
LU publication?
yes
id
db52fa93-cf21-4bcf-b5bc-cba83674cd4e
date added to LUP
2019-05-22 10:31:34
date last changed
2020-01-13 01:52:01
@article{db52fa93-cf21-4bcf-b5bc-cba83674cd4e,
  abstract     = {<p>In order to generate monoclonal antibodies (MAb) directed against the low molecular weight glycoprotein alpha 1-microglobulin, a BALB/c mouse was immunized with a mixture of human, guinea pig, rat and rabbit alpha 1-microglobulin homologues (multi-species immunization) and boosted several times. On day 194, the mouse splenocytes were fused to SP2/0 myeloma cells. The resulting hybridomas were screened for anti-alpha 1-microglobulin activity against the alpha 1-microglobulin mixture or against the individual homologues. For this screening, protein G (the newly described IgG-binding streptococcal protein) was used in a solid-phase radioimmunoassay. The binding of protein G to immobilized antigen-antibody complexes was enhanced by pre-incubation with rabbit anti-mouse immunoglobulin G. The result was a panel of nine established hybridoma lines, all producing unique monoclonal antibodies, of IgG1 or IgG2a class, to alpha 1-microglobulin. The antibodies were not only reactive in solid-phase radioimmunoassay, but they could also immunoprecipitate 125I-labeled soluble alpha 1-microglobulin. Moreover, they reacted specifically with the alpha 1-microglobulin band in Western blots of urinary proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Such monoclonal antibodies are potentially valuable reagents for the further characterization of alpha 1-microglobulin.</p>},
  author       = {Nilson, B and Akerström, B and Lögdberg, L},
  issn         = {0022-1759},
  language     = {eng},
  month        = {05},
  number       = {1},
  pages        = {39--45},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Cross-reacting monoclonal anti-alpha 1-microglobulin antibodies produced by multi-species immunization and using protein G for the screening assay},
  url          = {http://dx.doi.org/10.1016/0022-1759(87)90029-9},
  doi          = {10.1016/0022-1759(87)90029-9},
  volume       = {99},
  year         = {1987},
}