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Isolation and detection of human IgA using a streptococcal IgA-binding peptide.

Sandin, Charlotta LU ; Linse, Sara LU ; Areschoug, Thomas LU ; Woof, Jenny M; Reinholdt, Jesper and Lindahl, Gunnar LU (2002) In Journal of Immunology 169(3). p.1357-1364
Abstract
Bacterial proteins that bind to the Fc part of IgG have found widespread use in immunology. A similar protein suitable for the isolation and detection of human IgA has not been described. Here, we show that a 50-residue synthetic peptide, designated streptococcal IgA-binding peptide (Sap) and derived from a streptococcal M protein, can be used for single-step affinity purification of human IgA. High affinity binding of IgA required the presence in Sap of a C-terminal cysteine residue, not present in the intact M protein. Passage of human serum through a Sap column caused depletion of >99% of the IgA, and elution of the column allowed quantitative recovery of highly purified IgA, for which the proportions of the IgA1 and IgA2 subclasses... (More)
Bacterial proteins that bind to the Fc part of IgG have found widespread use in immunology. A similar protein suitable for the isolation and detection of human IgA has not been described. Here, we show that a 50-residue synthetic peptide, designated streptococcal IgA-binding peptide (Sap) and derived from a streptococcal M protein, can be used for single-step affinity purification of human IgA. High affinity binding of IgA required the presence in Sap of a C-terminal cysteine residue, not present in the intact M protein. Passage of human serum through a Sap column caused depletion of >99% of the IgA, and elution of the column allowed quantitative recovery of highly purified IgA, for which the proportions of the IgA1 and IgA2 subclasses were the same as in whole serum. Moreover, immobilized Sap could be used for single-step purification of secretory IgA of both subclasses from human saliva, with a recovery of approximately 45%. The Sap peptide could also be used to specifically detect IgA bound to Ag. Together, these data indicate that Sap is a versatile Fc-binding reagent that may open new possibilities for the characterization of human IgA. (Less)
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author
organization
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Contribution to journal
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published
subject
keywords
Carrier Proteins : metabolism, Rabbits, Molecular Sequence Data, Fc : metabolism, Immunoglobulins, Immunoglobulin A : metabolism, Immunoglobulin A : isolation & purification, Human, Immunoglobulin A : analysis, Bacterial Outer Membrane Proteins : metabolism, Animal, Amino Acid Sequence
in
Journal of Immunology
volume
169
issue
3
pages
1357 - 1364
publisher
American Association of Immunologists
external identifiers
  • wos:000177025100027
  • pmid:12133959
  • scopus:0036681978
ISSN
1550-6606
language
English
LU publication?
yes
id
77d04a3c-9e3a-41e0-900a-51b2138c2e46 (old id 109538)
alternative location
http://www.jimmunol.org/cgi/content/full/169/3/1357
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12133959&dopt=Abstract
date added to LUP
2007-07-10 13:00:10
date last changed
2017-11-12 04:03:11
@article{77d04a3c-9e3a-41e0-900a-51b2138c2e46,
  abstract     = {Bacterial proteins that bind to the Fc part of IgG have found widespread use in immunology. A similar protein suitable for the isolation and detection of human IgA has not been described. Here, we show that a 50-residue synthetic peptide, designated streptococcal IgA-binding peptide (Sap) and derived from a streptococcal M protein, can be used for single-step affinity purification of human IgA. High affinity binding of IgA required the presence in Sap of a C-terminal cysteine residue, not present in the intact M protein. Passage of human serum through a Sap column caused depletion of >99% of the IgA, and elution of the column allowed quantitative recovery of highly purified IgA, for which the proportions of the IgA1 and IgA2 subclasses were the same as in whole serum. Moreover, immobilized Sap could be used for single-step purification of secretory IgA of both subclasses from human saliva, with a recovery of approximately 45%. The Sap peptide could also be used to specifically detect IgA bound to Ag. Together, these data indicate that Sap is a versatile Fc-binding reagent that may open new possibilities for the characterization of human IgA.},
  author       = {Sandin, Charlotta and Linse, Sara and Areschoug, Thomas and Woof, Jenny M and Reinholdt, Jesper and Lindahl, Gunnar},
  issn         = {1550-6606},
  keyword      = {Carrier Proteins : metabolism,Rabbits,Molecular Sequence Data,Fc : metabolism,Immunoglobulins,Immunoglobulin A : metabolism,Immunoglobulin A : isolation & purification,Human,Immunoglobulin A : analysis,Bacterial Outer Membrane Proteins : metabolism,Animal,Amino Acid Sequence},
  language     = {eng},
  number       = {3},
  pages        = {1357--1364},
  publisher    = {American Association of Immunologists},
  series       = {Journal of Immunology},
  title        = {Isolation and detection of human IgA using a streptococcal IgA-binding peptide.},
  volume       = {169},
  year         = {2002},
}