The impact of impurities in synthetic peptides on the outcome of T‐cell stimulation assays
Identifieur interne : 002C20 ( Main/Exploration ); précédent : 002C19; suivant : 002C21The impact of impurities in synthetic peptides on the outcome of T‐cell stimulation assays
Auteurs : Janet W. De Beukelaar [Pays-Bas] ; Jan W. Gratama [Pays-Bas] ; Peter A. Sillevis Smitt [Pays-Bas] ; Georges M. Verjans [Pays-Bas] ; Jaco Kraan [Pays-Bas] ; Theo M. Luider [Pays-Bas] ; Peter C. Burgers [Pays-Bas]Source :
- Rapid Communications in Mass Spectrometry [ 0951-4198 ] ; 2007-04-15.
English descriptors
- Teeft :
- Amino, Amino acid, Amino acids, Assay, Assay outcome, Base peak, Base peak protonated peptide, Batch, Charge delocalization, Commun, Comparative analysis, Complete series, Copyright, Deletion, Deletion peptide, Deletion peptides, Different batches, Different purities, Erasmus university, Exact mass, Exact mass measurements, Exact mass measurements show, External calibration, Fragmentation conditions, Ftms, Ftms spectra, Impurity, Isotope peak, John wiley sons, Liquid chromatography, Mass spectra, Mass spectrom, Mass spectrometry, Mass spectrum, Negative control, Netherlands proteomics centre, Numerous differences, Partial ftms spectrum, Peptide, Peptide batches, Peptide impurity, Peptide pools, Peptide synthesis, Positive responses, Product ions, Protonated, Protonated peptide, Purity batch, Purity peptides, Rapid commun, Same manufacturer, Same peptide, Sequence gftmtnydeaamai, Spectrom, Strong calcium, Such experiments, Sulfated, Sulfated peptide, Sulfated peptides, Synthesis method, Synthetic peptide, Synthetic peptides, Target plate, Wide range.
Abstract
Protein‐spanning peptide pools have proven valuable as a screening tool for detecting T‐lymphocyte responses against a wide range of proteins. We have used this approach in our search for T cells reactive to the onconeural protein HuD. We found positive responses in only 3 of 127 individuals; however, these were highly unusual in that the same class I HLA alleles and peptides were involved. These T‐cell responses were not confirmed when peptides re‐synthesized by the same manufacturer with similar and with higher purity levels were used. Our observations indicated that these T‐cell responses were not directed against the designed HuD peptides. Here, we report on (i) comparisons of the peptide batches analyzed by matrix‐assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI‐FTMS) that did –and did not – elicit T‐cell responses and (ii) a detailed analysis of the various by‐products of peptides, irrespective of T‐cell assay outcome. We found numerous differences between the peptide batches, such as omissions of amino acids in the primary structure of the peptides. Furthermore, some batches revealed strong interactions with calcium ions or contained sulfated peptides. Our data reveal that different batches from the same peptide may contain artefacts that influence the outcome of HLA‐restricted T‐cell response assays. Copyright © 2007 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/rcm.2958
Affiliations:
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De Beukelaar</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Laboratory of Neuro‐Oncology, Department of Neurology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Medical Oncology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Gratama, Jan W" sort="Gratama, Jan W" uniqKey="Gratama J" first="Jan W." last="Gratama">Jan W. Gratama</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Medical Oncology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Smitt, Peter A Sillevis" sort="Smitt, Peter A Sillevis" uniqKey="Smitt P" first="Peter A. Sillevis" last="Smitt">Peter A. Sillevis Smitt</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Laboratory of Neuro‐Oncology, Department of Neurology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Verjans, Georges M" sort="Verjans, Georges M" uniqKey="Verjans G" first="Georges M." last="Verjans">Georges M. Verjans</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Virology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Kraan, Jaco" sort="Kraan, Jaco" uniqKey="Kraan J" first="Jaco" last="Kraan">Jaco Kraan</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Medical Oncology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Luider, Theo M" sort="Luider, Theo M" uniqKey="Luider T" first="Theo M." last="Luider">Theo M. Luider</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Laboratory of Neuro‐Oncology, Department of Neurology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C." last="Burgers">Peter C. Burgers</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Laboratory of Neuro‐Oncology, Department of Neurology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Pays-Bas</country></affiliation><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Correspondence address: Laboratory of Neuro‐Oncology, Department of Neurology Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Rapid Communications in Mass Spectrometry</title><title level="j" type="alt">RAPID COMMUNICATIONS IN MASS SPECTROMETRY</title><idno type="ISSN">0951-4198</idno><idno type="eISSN">1097-0231</idno><imprint><biblScope unit="vol">21</biblScope><biblScope unit="issue">7</biblScope><biblScope unit="page" from="1282">1282</biblScope><biblScope unit="page" to="1288">1288</biblScope><biblScope unit="page-count">7</biblScope><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2007-04-15">2007-04-15</date></imprint><idno type="ISSN">0951-4198</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0951-4198</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Amino</term><term>Amino acid</term><term>Amino acids</term><term>Assay</term><term>Assay outcome</term><term>Base peak</term><term>Base peak protonated peptide</term><term>Batch</term><term>Charge delocalization</term><term>Commun</term><term>Comparative analysis</term><term>Complete series</term><term>Copyright</term><term>Deletion</term><term>Deletion peptide</term><term>Deletion peptides</term><term>Different batches</term><term>Different purities</term><term>Erasmus university</term><term>Exact mass</term><term>Exact mass measurements</term><term>Exact mass measurements show</term><term>External calibration</term><term>Fragmentation conditions</term><term>Ftms</term><term>Ftms spectra</term><term>Impurity</term><term>Isotope peak</term><term>John wiley sons</term><term>Liquid chromatography</term><term>Mass spectra</term><term>Mass spectrom</term><term>Mass spectrometry</term><term>Mass spectrum</term><term>Negative control</term><term>Netherlands proteomics centre</term><term>Numerous differences</term><term>Partial ftms spectrum</term><term>Peptide</term><term>Peptide batches</term><term>Peptide impurity</term><term>Peptide pools</term><term>Peptide synthesis</term><term>Positive responses</term><term>Product ions</term><term>Protonated</term><term>Protonated peptide</term><term>Purity batch</term><term>Purity peptides</term><term>Rapid commun</term><term>Same manufacturer</term><term>Same peptide</term><term>Sequence gftmtnydeaamai</term><term>Spectrom</term><term>Strong calcium</term><term>Such experiments</term><term>Sulfated</term><term>Sulfated peptide</term><term>Sulfated peptides</term><term>Synthesis method</term><term>Synthetic peptide</term><term>Synthetic peptides</term><term>Target plate</term><term>Wide range</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Protein‐spanning peptide pools have proven valuable as a screening tool for detecting T‐lymphocyte responses against a wide range of proteins. We have used this approach in our search for T cells reactive to the onconeural protein HuD. We found positive responses in only 3 of 127 individuals; however, these were highly unusual in that the same class I HLA alleles and peptides were involved. These T‐cell responses were not confirmed when peptides re‐synthesized by the same manufacturer with similar and with higher purity levels were used. Our observations indicated that these T‐cell responses were not directed against the designed HuD peptides. Here, we report on (i) comparisons of the peptide batches analyzed by matrix‐assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI‐FTMS) that did –and did not – elicit T‐cell responses and (ii) a detailed analysis of the various by‐products of peptides, irrespective of T‐cell assay outcome. We found numerous differences between the peptide batches, such as omissions of amino acids in the primary structure of the peptides. Furthermore, some batches revealed strong interactions with calcium ions or contained sulfated peptides. Our data reveal that different batches from the same peptide may contain artefacts that influence the outcome of HLA‐restricted T‐cell response assays. Copyright © 2007 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>Pays-Bas</li></country><region><li>Hollande-Méridionale</li></region><settlement><li>Rotterdam</li></settlement></list><tree><country name="Pays-Bas"><region name="Hollande-Méridionale"><name sortKey="De Beukelaar, Janet W" sort="De Beukelaar, Janet W" uniqKey="De Beukelaar J" first="Janet W." last="De Beukelaar">Janet W. De Beukelaar</name></region><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C." last="Burgers">Peter C. Burgers</name><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C." last="Burgers">Peter C. Burgers</name><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C." last="Burgers">Peter C. Burgers</name><name sortKey="De Beukelaar, Janet W" sort="De Beukelaar, Janet W" uniqKey="De Beukelaar J" first="Janet W." last="De Beukelaar">Janet W. De Beukelaar</name><name sortKey="Gratama, Jan W" sort="Gratama, Jan W" uniqKey="Gratama J" first="Jan W." last="Gratama">Jan W. Gratama</name><name sortKey="Kraan, Jaco" sort="Kraan, Jaco" uniqKey="Kraan J" first="Jaco" last="Kraan">Jaco Kraan</name><name sortKey="Luider, Theo M" sort="Luider, Theo M" uniqKey="Luider T" first="Theo M." last="Luider">Theo M. Luider</name><name sortKey="Smitt, Peter A Sillevis" sort="Smitt, Peter A Sillevis" uniqKey="Smitt P" first="Peter A. Sillevis" last="Smitt">Peter A. Sillevis Smitt</name><name sortKey="Verjans, Georges M" sort="Verjans, Georges M" uniqKey="Verjans G" first="Georges M." last="Verjans">Georges M. 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