Purification of antibodies using protein L-binding framework structures in the light chain variable domain
(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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- author
- Nilson, B H LU ; Lögdberg, L ; Kastern, W ; Björck, L LU and Akerström, B LU
- organization
- publishing date
- 1993-08-26
- 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
- 2024-08-06 18:33:20
@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}}, }