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Antibody response in immunized rabbits measured with bacterial immunoglobulin-binding proteins

Elbashir, M I ; Nilson, B H LU orcid ; Akesson, P ; Björck, L LU and Akerström, B LU (1990) In Journal of Immunological Methods 135(1-2). p.9-171
Abstract

Protein G, an immunoglobulin (Ig)-binding protein isolated from group C or G streptococci, binds to the Fc portion of IgG. Protein L, from the anaerobic bacterium Peptostreptococcus magnus, specifically binds light chains of Ig. In this study, protein G and L were used to measure the production of antibodies in immunized rabbits. Two rabbits were immunized with a mixture of human urinary proteins from a patient with tubular proteinuria, and blood samples were collected regularly from the animals for 6 weeks after the immunization. The antibody levels of the blood samples against six of the proteins in the antigen mixture were then measured by ELISA. Microtiter plates were coated with each of the antigens, incubated with the rabbit serum... (More)

Protein G, an immunoglobulin (Ig)-binding protein isolated from group C or G streptococci, binds to the Fc portion of IgG. Protein L, from the anaerobic bacterium Peptostreptococcus magnus, specifically binds light chains of Ig. In this study, protein G and L were used to measure the production of antibodies in immunized rabbits. Two rabbits were immunized with a mixture of human urinary proteins from a patient with tubular proteinuria, and blood samples were collected regularly from the animals for 6 weeks after the immunization. The antibody levels of the blood samples against six of the proteins in the antigen mixture were then measured by ELISA. Microtiter plates were coated with each of the antigens, incubated with the rabbit serum samples, and the specific antibodies of the IgG class measured by incubation with biotinylated protein G, and antibodies of all Ig classes with biotinylated protein L. Alternatively, Western blotting was employed, where the antibodies which bound to each antigen after separation by SDS-PAGE and transfer to nitrocellulose membranes, were detected by protein G or L. The results showed that antibody production against five of the antigens, albumin, alpha 1 gamma-acid glycoprotein, alpha 1 gamma-microglobulin, Ig light chains, and retinol-binding protein, showed a similar pattern, although the magnitude of the initial IgM response differed somewhat. After 6 weeks, the levels of the protein G-binding antibodies had reached a plateau, while those of protein L-binding antibodies were still increasing. The response to the sixth antigen, beta 2 microglobulin, was considerably different. A dramatic increase of anti-beta 2 gamma-microglobulin antibodies was seen during the 4th week after immunization when protein L was used.

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author
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organization
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Contribution to journal
publication status
published
subject
keywords
Animals, Antibody Formation, Bacterial Proteins, Biotin, Blotting, Western, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Immunization, Immunoglobulin Fc Fragments, Immunologic Techniques, Proteins, Proteinuria, Rabbits, Journal Article, Research Support, Non-U.S. Gov't
in
Journal of Immunological Methods
volume
135
issue
1-2
pages
9 - 171
publisher
Elsevier
external identifiers
  • scopus:0025663451
  • pmid:2273256
ISSN
0022-1759
DOI
10.1016/0022-1759(90)90270-6
language
English
LU publication?
yes
id
c9cea130-9870-46fa-9a1c-7a0653dd26f0
date added to LUP
2018-05-26 14:09:20
date last changed
2024-03-18 08:19:36
@article{c9cea130-9870-46fa-9a1c-7a0653dd26f0,
  abstract     = {{<p>Protein G, an immunoglobulin (Ig)-binding protein isolated from group C or G streptococci, binds to the Fc portion of IgG. Protein L, from the anaerobic bacterium Peptostreptococcus magnus, specifically binds light chains of Ig. In this study, protein G and L were used to measure the production of antibodies in immunized rabbits. Two rabbits were immunized with a mixture of human urinary proteins from a patient with tubular proteinuria, and blood samples were collected regularly from the animals for 6 weeks after the immunization. The antibody levels of the blood samples against six of the proteins in the antigen mixture were then measured by ELISA. Microtiter plates were coated with each of the antigens, incubated with the rabbit serum samples, and the specific antibodies of the IgG class measured by incubation with biotinylated protein G, and antibodies of all Ig classes with biotinylated protein L. Alternatively, Western blotting was employed, where the antibodies which bound to each antigen after separation by SDS-PAGE and transfer to nitrocellulose membranes, were detected by protein G or L. The results showed that antibody production against five of the antigens, albumin, alpha 1 gamma-acid glycoprotein, alpha 1 gamma-microglobulin, Ig light chains, and retinol-binding protein, showed a similar pattern, although the magnitude of the initial IgM response differed somewhat. After 6 weeks, the levels of the protein G-binding antibodies had reached a plateau, while those of protein L-binding antibodies were still increasing. The response to the sixth antigen, beta 2 microglobulin, was considerably different. A dramatic increase of anti-beta 2 gamma-microglobulin antibodies was seen during the 4th week after immunization when protein L was used.</p>}},
  author       = {{Elbashir, M I and Nilson, B H and Akesson, P and Björck, L and Akerström, B}},
  issn         = {{0022-1759}},
  keywords     = {{Animals; Antibody Formation; Bacterial Proteins; Biotin; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Immunization; Immunoglobulin Fc Fragments; Immunologic Techniques; Proteins; Proteinuria; Rabbits; Journal Article; Research Support, Non-U.S. Gov't}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{1-2}},
  pages        = {{9--171}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Immunological Methods}},
  title        = {{Antibody response in immunized rabbits measured with bacterial immunoglobulin-binding proteins}},
  url          = {{http://dx.doi.org/10.1016/0022-1759(90)90270-6}},
  doi          = {{10.1016/0022-1759(90)90270-6}},
  volume       = {{135}},
  year         = {{1990}},
}