Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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

Nilson, B H LU orcid ; 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.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; and
organization
publishing date
type
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
  • pmid:8360508
  • 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
2025-10-15 21:33:20
@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}},
  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}},
  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}},
  doi          = {{10.1016/0022-1759(93)90273-A}},
  volume       = {{164}},
  year         = {{1993}},
}