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.
(Less)
- author
- Nilson, B
LU
; Akerström, B
LU
and Lögdberg, L
- organization
- publishing date
- 1987-05-04
- 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
- 2024-01-01 06:54:41
@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}},
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}},
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}},
}