Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3.
Mass spectrometry (MS) analysis for detection of immunoglobulins (IG) of the human IgG3 subclass is described that relies on polymorphic amino acids of the heavy gamma3 chains. IgG3 is the most polymorphic human IgG subclass with thirteen G3m allotypes located on the constant CH2 and CH3 domains of...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3457951?pdf=render |
| _version_ | 1819265805009813504 |
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| author | Célia Dechavanne François Guillonneau Giovanni Chiappetta Laïla Sago Prisca Lévy Virginie Salnot Evelyne Guitard François Ehrenmann Cédric Broussard Philippe Chafey Agnès Le Port Joëlle Vinh Patrick Mayeux Jean-Michel Dugoujon Marie-Paule Lefranc Florence Migot-Nabias |
| author_facet | Célia Dechavanne François Guillonneau Giovanni Chiappetta Laïla Sago Prisca Lévy Virginie Salnot Evelyne Guitard François Ehrenmann Cédric Broussard Philippe Chafey Agnès Le Port Joëlle Vinh Patrick Mayeux Jean-Michel Dugoujon Marie-Paule Lefranc Florence Migot-Nabias |
| author_sort | Célia Dechavanne |
| collection | DOAJ |
| description | Mass spectrometry (MS) analysis for detection of immunoglobulins (IG) of the human IgG3 subclass is described that relies on polymorphic amino acids of the heavy gamma3 chains. IgG3 is the most polymorphic human IgG subclass with thirteen G3m allotypes located on the constant CH2 and CH3 domains of the gamma3 chain, the combination of which leads to six major G3m alleles. Amino acid changes resulting of extensive sequencing previously led to the definition of 19 IGHG3 alleles that have been correlated to the G3m alleles. As a proof of concept, MS proteotypic peptides were defined which encompass discriminatory amino acids for the identification of the G3m and IGHG3 alleles. Plasma samples originating from ten individuals either homozygous or heterozygous for different G3m alleles, and including one mother and her baby (drawn sequentially from birth to 9 months of age), were analyzed. Total IgG3 were purified using affinity chromatography and then digested by a combination of AspN and trypsin proteases, and peptides of interest were detected by mass spectrometry. The sensitivity of the method was assessed by mixing variable amounts of two plasma samples bearing distinct G3m allotypes. A label-free approach using the high-performance liquid chromatography (HPLC) retention time of peptides and their MS mass analyzer peak intensity gave semi-quantitative information. Quantification was realized by selected reaction monitoring (SRM) using synthetic peptides as internal standards. The possibility offered by this new methodology to detect and quantify neo-synthesized IgG in newborns will improve knowledge on the first acquisition of antibodies in infants and constitutes a promising diagnostic tool for vertically-transmitted diseases. |
| first_indexed | 2024-12-23T20:51:12Z |
| format | Article |
| id | doaj.art-c0988dab377946eea127195753cc8c4e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T20:51:12Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-c0988dab377946eea127195753cc8c4e2022-12-21T17:31:39ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0179e4609710.1371/journal.pone.0046097Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3.Célia DechavanneFrançois GuillonneauGiovanni ChiappettaLaïla SagoPrisca LévyVirginie SalnotEvelyne GuitardFrançois EhrenmannCédric BroussardPhilippe ChafeyAgnès Le PortJoëlle VinhPatrick MayeuxJean-Michel DugoujonMarie-Paule LefrancFlorence Migot-NabiasMass spectrometry (MS) analysis for detection of immunoglobulins (IG) of the human IgG3 subclass is described that relies on polymorphic amino acids of the heavy gamma3 chains. IgG3 is the most polymorphic human IgG subclass with thirteen G3m allotypes located on the constant CH2 and CH3 domains of the gamma3 chain, the combination of which leads to six major G3m alleles. Amino acid changes resulting of extensive sequencing previously led to the definition of 19 IGHG3 alleles that have been correlated to the G3m alleles. As a proof of concept, MS proteotypic peptides were defined which encompass discriminatory amino acids for the identification of the G3m and IGHG3 alleles. Plasma samples originating from ten individuals either homozygous or heterozygous for different G3m alleles, and including one mother and her baby (drawn sequentially from birth to 9 months of age), were analyzed. Total IgG3 were purified using affinity chromatography and then digested by a combination of AspN and trypsin proteases, and peptides of interest were detected by mass spectrometry. The sensitivity of the method was assessed by mixing variable amounts of two plasma samples bearing distinct G3m allotypes. A label-free approach using the high-performance liquid chromatography (HPLC) retention time of peptides and their MS mass analyzer peak intensity gave semi-quantitative information. Quantification was realized by selected reaction monitoring (SRM) using synthetic peptides as internal standards. The possibility offered by this new methodology to detect and quantify neo-synthesized IgG in newborns will improve knowledge on the first acquisition of antibodies in infants and constitutes a promising diagnostic tool for vertically-transmitted diseases.http://europepmc.org/articles/PMC3457951?pdf=render |
| spellingShingle | Célia Dechavanne François Guillonneau Giovanni Chiappetta Laïla Sago Prisca Lévy Virginie Salnot Evelyne Guitard François Ehrenmann Cédric Broussard Philippe Chafey Agnès Le Port Joëlle Vinh Patrick Mayeux Jean-Michel Dugoujon Marie-Paule Lefranc Florence Migot-Nabias Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. PLoS ONE |
| title | Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. |
| title_full | Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. |
| title_fullStr | Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. |
| title_full_unstemmed | Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. |
| title_short | Mass spectrometry detection of G3m and IGHG3 alleles and follow-up of differential mother and neonate IgG3. |
| title_sort | mass spectrometry detection of g3m and ighg3 alleles and follow up of differential mother and neonate igg3 |
| url | http://europepmc.org/articles/PMC3457951?pdf=render |
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