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Purification of antibodies using protein L-binding framework structures in the light chain variable domain

Nilson, B H LU ; Lögdberg, L; Kastern, W; Björck, L LU and Akerström, B LU (1993) In Journal of Immunological Methods 164(1). p.33-40
Abstract

Protein L from the bacterial species Peptostreptococcus magnus binds specifically to the variable domain of Ig light chains, without interfering with the antigen-binding site. In this work a genetically engineered fragment of protein L, including four of the repeated Ig-binding repeat units, was employed for the purification of Ig from various sources. Thus, IgG, IgM, and IgA were purified from human and mouse serum in a single step using protein L-Sepharose affinity chromatography. Moreover, human and mouse monoclonal IgG, IgM, and IgA, and human IgG Fab fragments, as well as a mouse/human chimeric recombinant antibody, could be purified from cultures of hybridoma cells or antibody-producing bacterial cells, with protein L-Sepharose.... (More)

Protein L from the bacterial species Peptostreptococcus magnus binds specifically to the variable domain of Ig light chains, without interfering with the antigen-binding site. In this work a genetically engineered fragment of protein L, including four of the repeated Ig-binding repeat units, was employed for the purification of Ig from various sources. Thus, IgG, IgM, and IgA were purified from human and mouse serum in a single step using protein L-Sepharose affinity chromatography. Moreover, human and mouse monoclonal IgG, IgM, and IgA, and human IgG Fab fragments, as well as a mouse/human chimeric recombinant antibody, could be purified from cultures of hybridoma cells or antibody-producing bacterial cells, with protein L-Sepharose. This was also the case with a humanized mouse antibody, in which mouse hypervariable antigen-binding regions had been introduced into a protein L-binding kappa subtype III human IgG. These experiments demonstrate that it is possible to engineer antibodies and antibody fragments (Fab, Fv) with protein L-binding framework regions, which can then be utilized in a protein L-based purification protocol.

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organization
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Contribution to journal
publication status
published
subject
keywords
Animals, Antibodies, Bacterial Proteins, Chromatography, Affinity, Humans, Immunoglobulin Light Chains, Mice, Papio, Peptostreptococcus, Recombinant Fusion Proteins, Journal Article, Research Support, Non-U.S. Gov't
in
Journal of Immunological Methods
volume
164
issue
1
pages
33 - 40
publisher
Elsevier
external identifiers
  • scopus:0027249218
ISSN
0022-1759
DOI
10.1016/0022-1759(93)90273-A
language
English
LU publication?
yes
id
dad3c1a9-80a5-4c9c-ba9b-d11f5c18ac8f
date added to LUP
2018-05-26 14:02:42
date last changed
2018-10-03 11:38:52
@article{dad3c1a9-80a5-4c9c-ba9b-d11f5c18ac8f,
  abstract     = {<p>Protein L from the bacterial species Peptostreptococcus magnus binds specifically to the variable domain of Ig light chains, without interfering with the antigen-binding site. In this work a genetically engineered fragment of protein L, including four of the repeated Ig-binding repeat units, was employed for the purification of Ig from various sources. Thus, IgG, IgM, and IgA were purified from human and mouse serum in a single step using protein L-Sepharose affinity chromatography. Moreover, human and mouse monoclonal IgG, IgM, and IgA, and human IgG Fab fragments, as well as a mouse/human chimeric recombinant antibody, could be purified from cultures of hybridoma cells or antibody-producing bacterial cells, with protein L-Sepharose. This was also the case with a humanized mouse antibody, in which mouse hypervariable antigen-binding regions had been introduced into a protein L-binding kappa subtype III human IgG. These experiments demonstrate that it is possible to engineer antibodies and antibody fragments (Fab, Fv) with protein L-binding framework regions, which can then be utilized in a protein L-based purification protocol.</p>},
  author       = {Nilson, B H and Lögdberg, L and Kastern, W and Björck, L and Akerström, B},
  issn         = {0022-1759},
  keyword      = {Animals,Antibodies,Bacterial Proteins,Chromatography, Affinity,Humans,Immunoglobulin Light Chains,Mice,Papio,Peptostreptococcus,Recombinant Fusion Proteins,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  month        = {08},
  number       = {1},
  pages        = {33--40},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Purification of antibodies using protein L-binding framework structures in the light chain variable domain},
  url          = {http://dx.doi.org/10.1016/0022-1759(93)90273-A},
  volume       = {164},
  year         = {1993},
}