Identification and Mapping of Linear Antibody Epitopes in Human Serum Albumin Using High-Density Peptide Arrays
Identifieur interne : 000A98 ( Ncbi/Merge ); précédent : 000A97; suivant : 000A99Identification and Mapping of Linear Antibody Epitopes in Human Serum Albumin Using High-Density Peptide Arrays
Auteurs : Lajla Bruntse Hansen [Danemark] ; Soren Buus [Danemark] ; Claus Schafer-Nielsen [Danemark]Source :
- PLoS ONE [ 1932-6203 ] ; 2013.
Descripteurs français
- KwdFr :
- Adulte d'âge moyen, Animaux, Bovins, Cartographie cérébrale, Dysfonctionnement cognitif (diagnostic), Dysfonctionnement cognitif (métabolisme), Femelle, Fluorodésoxyglucose F18, Glucose (métabolisme), Humains, Lapins, Maladie d'Alzheimer (diagnostic), Maladie d'Alzheimer (métabolisme), Mâle, Neuroimagerie, Phénomènes physiologiques cellulaires, Sujet âgé, Sujet âgé de 80 ans ou plus, Tomographie par émission de positons, Études cas-témoins, Études de suivi, Études longitudinales.
- MESH :
- diagnostic : Dysfonctionnement cognitif, Maladie d'Alzheimer.
- métabolisme : Dysfonctionnement cognitif, Glucose, Maladie d'Alzheimer.
- Adulte d'âge moyen, Animaux, Bovins, Cartographie cérébrale, Femelle, Fluorodésoxyglucose F18, Humains, Lapins, Mâle, Neuroimagerie, Phénomènes physiologiques cellulaires, Sujet âgé, Sujet âgé de 80 ans ou plus, Tomographie par émission de positons, Études cas-témoins, Études de suivi, Études longitudinales.
English descriptors
- KwdEn :
- Aged, Aged, 80 and over, Alzheimer Disease (diagnosis), Alzheimer Disease (metabolism), Animals, Brain Mapping, Case-Control Studies, Cattle, Cell Physiological Phenomena, Cognitive Dysfunction (diagnosis), Cognitive Dysfunction (metabolism), Female, Fluorodeoxyglucose F18, Follow-Up Studies, Glucose (metabolism), Humans, Longitudinal Studies, Male, Middle Aged, Neural Networks, Computer, Neuroimaging, Positron-Emission Tomography, Rabbits.
- MESH :
- chemical , metabolism : Glucose.
- chemical : Fluorodeoxyglucose F18.
- diagnosis : Alzheimer Disease, Cognitive Dysfunction.
- metabolism : Alzheimer Disease, Cognitive Dysfunction.
- Aged, Aged, 80 and over, Animals, Brain Mapping, Case-Control Studies, Cattle, Cell Physiological Phenomena, Female, Follow-Up Studies, Humans, Longitudinal Studies, Male, Middle Aged, Neural Networks, Computer, Neuroimaging, Positron-Emission Tomography, Rabbits.
Abstract
We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm2. Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.
Url:
DOI: 10.1371/journal.pone.0068902
PubMed: 23894373
PubMed Central: 3720873
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PMC:3720873Le document en format XML
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wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Schafer-N, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Schafer-N, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Buus, Soren" sort="Buus, Soren" uniqKey="Buus S" first="Soren" last="Buus">Soren Buus</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental 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type="eISSN">1932-6203</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aged</term><term>Aged, 80 and over</term><term>Alzheimer Disease (diagnosis)</term><term>Alzheimer Disease (metabolism)</term><term>Animals</term><term>Brain Mapping</term><term>Case-Control Studies</term><term>Cattle</term><term>Cell Physiological Phenomena</term><term>Cognitive Dysfunction (diagnosis)</term><term>Cognitive Dysfunction (metabolism)</term><term>Female</term><term>Fluorodeoxyglucose F18</term><term>Follow-Up Studies</term><term>Glucose (metabolism)</term><term>Humans</term><term>Longitudinal Studies</term><term>Male</term><term>Middle Aged</term><term>Neural Networks, Computer</term><term>Neuroimaging</term><term>Positron-Emission Tomography</term><term>Rabbits</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adulte d'âge moyen</term><term>Animaux</term><term>Bovins</term><term>Cartographie cérébrale</term><term>Dysfonctionnement cognitif (diagnostic)</term><term>Dysfonctionnement cognitif (métabolisme)</term><term>Femelle</term><term>Fluorodésoxyglucose F18</term><term>Glucose (métabolisme)</term><term>Humains</term><term>Lapins</term><term>Maladie d'Alzheimer (diagnostic)</term><term>Maladie d'Alzheimer (métabolisme)</term><term>Mâle</term><term>Neuroimagerie</term><term>Phénomènes physiologiques cellulaires</term><term>Sujet âgé</term><term>Sujet âgé de 80 ans ou plus</term><term>Tomographie par émission de positons</term><term>Études cas-témoins</term><term>Études de suivi</term><term>Études longitudinales</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glucose</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Fluorodeoxyglucose F18</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Alzheimer Disease</term><term>Cognitive Dysfunction</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Dysfonctionnement cognitif</term><term>Maladie d'Alzheimer</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Alzheimer Disease</term><term>Cognitive Dysfunction</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Dysfonctionnement cognitif</term><term>Glucose</term><term>Maladie d'Alzheimer</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Aged</term><term>Aged, 80 and over</term><term>Animals</term><term>Brain Mapping</term><term>Case-Control Studies</term><term>Cattle</term><term>Cell Physiological Phenomena</term><term>Female</term><term>Follow-Up Studies</term><term>Humans</term><term>Longitudinal Studies</term><term>Male</term><term>Middle Aged</term><term>Neural Networks, Computer</term><term>Neuroimaging</term><term>Positron-Emission Tomography</term><term>Rabbits</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adulte d'âge moyen</term><term>Animaux</term><term>Bovins</term><term>Cartographie cérébrale</term><term>Femelle</term><term>Fluorodésoxyglucose F18</term><term>Humains</term><term>Lapins</term><term>Mâle</term><term>Neuroimagerie</term><term>Phénomènes physiologiques cellulaires</term><term>Sujet âgé</term><term>Sujet âgé de 80 ans ou plus</term><term>Tomographie par émission de positons</term><term>Études cas-témoins</term><term>Études de suivi</term><term>Études longitudinales</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm<sup>2</sup>. Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Saul, Fa" uniqKey="Saul F">FA Saul</name></author><author><name sortKey="Alzari, Pm" uniqKey="Alzari P">PM Alzari</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, Hl" uniqKey="Liu H">HL Liu</name></author><author><name sortKey="Hsu, Jp" uniqKey="Hsu J">JP Hsu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Goetze, Am" uniqKey="Goetze A">AM Goetze</name></author><author><name sortKey="Richards, Jh" uniqKey="Richards J">JH Richards</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rosen, O" uniqKey="Rosen O">O Rosen</name></author><author><name sortKey="Anglister, J" uniqKey="Anglister J">J Anglister</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dhungana, S" uniqKey="Dhungana S">S Dhungana</name></author><author><name sortKey="Williams, Jg" uniqKey="Williams J">JG Williams</name></author><author><name sortKey="Fessler, Mb" uniqKey="Fessler M">MB Fessler</name></author><author><name sortKey="Tomer, Kb" uniqKey="Tomer K">KB Tomer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cretich, M" uniqKey="Cretich M">M Cretich</name></author><author><name sortKey="Damin, F" uniqKey="Damin F">F Damin</name></author><author><name sortKey="Pirri, G" uniqKey="Pirri G">G Pirri</name></author><author><name sortKey="Chiari, M" uniqKey="Chiari M">M Chiari</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chandra, H" uniqKey="Chandra H">H Chandra</name></author><author><name sortKey="Srivastava, S" uniqKey="Srivastava S">S Srivastava</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Benjamin, Dc" uniqKey="Benjamin D">DC Benjamin</name></author><author><name sortKey="Perdue, Ss" uniqKey="Perdue S">SS Perdue</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pande, J" uniqKey="Pande J">J Pande</name></author><author><name sortKey="Szewczyk, Mm" uniqKey="Szewczyk M">MM Szewczyk</name></author><author><name sortKey="Grover, Ak" uniqKey="Grover A">AK Grover</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Timmerman, P" uniqKey="Timmerman P">P Timmerman</name></author><author><name sortKey="Puijk, Wc" uniqKey="Puijk W">WC Puijk</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Van Regenmortel, Mh" uniqKey="Van Regenmortel M">MH Van Regenmortel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Geysen, Hm" uniqKey="Geysen H">HM Geysen</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author><author><name sortKey="Barteling, Sj" uniqKey="Barteling S">SJ Barteling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Houghten, Ra" uniqKey="Houghten R">RA Houghten</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Furka, A" uniqKey="Furka A">A Furka</name></author><author><name sortKey="Sebestyen, F" uniqKey="Sebestyen F">F Sebestyen</name></author><author><name sortKey="Asgedom, M" uniqKey="Asgedom M">M Asgedom</name></author><author><name sortKey="Dibo, G" uniqKey="Dibo G">G Dibo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lam, Ks" uniqKey="Lam K">KS Lam</name></author><author><name sortKey="Salmon, Se" uniqKey="Salmon S">SE 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sortKey="Forsstrom, B" uniqKey="Forsstrom B">B Forsstrom</name></author><author><name sortKey="Nilsson, P" uniqKey="Nilsson P">P Nilsson</name></author><author><name sortKey="Uhlen, M" uniqKey="Uhlen M">M Uhlen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yxfeldt, G" uniqKey="Yxfeldt G">G Yxfeldt</name></author><author><name sortKey="Froman, G" uniqKey="Froman G">G Froman</name></author><author><name sortKey="Mardh, Pa" uniqKey="Mardh P">PA Mardh</name></author><author><name sortKey="Ward, Me" uniqKey="Ward M">ME Ward</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Timmerman, P" uniqKey="Timmerman P">P Timmerman</name></author><author><name sortKey="Beld, J" uniqKey="Beld J">J Beld</name></author><author><name sortKey="Puijk, Wc" uniqKey="Puijk W">WC Puijk</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author></analytic></biblStruct></listBibl></div1></back></TEI><double pmid="23894373"><pmc><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Identification and Mapping of Linear Antibody Epitopes in Human Serum Albumin Using High-Density Peptide Arrays</title><author><name sortKey="Hansen, Lajla Bruntse" sort="Hansen, Lajla Bruntse" uniqKey="Hansen L" first="Lajla Bruntse" last="Hansen">Lajla Bruntse Hansen</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Schafer-N, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Schafer-N, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Buus, Soren" sort="Buus, Soren" uniqKey="Buus S" first="Soren" last="Buus">Soren Buus</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Schafer-N, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Schafer-N, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23894373</idno><idno type="pmc">3720873</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3720873</idno><idno type="RBID">PMC:3720873</idno><idno type="doi">10.1371/journal.pone.0068902</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">001080</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001080</idno><idno type="wicri:Area/Pmc/Curation">001080</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001080</idno><idno type="wicri:Area/Pmc/Checkpoint">001220</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001220</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Identification and Mapping of Linear Antibody Epitopes in Human Serum Albumin Using High-Density Peptide Arrays</title><author><name sortKey="Hansen, Lajla Bruntse" sort="Hansen, Lajla Bruntse" uniqKey="Hansen L" first="Lajla Bruntse" last="Hansen">Lajla Bruntse Hansen</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Schafer-N, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Schafer-N, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Buus, Soren" sort="Buus, Soren" uniqKey="Buus S" first="Soren" last="Buus">Soren Buus</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Schafer-N, Copenhagen, Denmark</addr-line></nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Schafer-N, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm<sup>2</sup>. Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Saul, Fa" uniqKey="Saul F">FA Saul</name></author><author><name sortKey="Alzari, Pm" uniqKey="Alzari P">PM Alzari</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, Hl" uniqKey="Liu H">HL Liu</name></author><author><name sortKey="Hsu, Jp" uniqKey="Hsu J">JP Hsu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Goetze, Am" uniqKey="Goetze A">AM Goetze</name></author><author><name sortKey="Richards, Jh" uniqKey="Richards J">JH Richards</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rosen, O" uniqKey="Rosen O">O Rosen</name></author><author><name sortKey="Anglister, J" uniqKey="Anglister J">J Anglister</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dhungana, S" uniqKey="Dhungana S">S Dhungana</name></author><author><name sortKey="Williams, Jg" uniqKey="Williams J">JG Williams</name></author><author><name sortKey="Fessler, Mb" uniqKey="Fessler M">MB Fessler</name></author><author><name sortKey="Tomer, Kb" uniqKey="Tomer K">KB Tomer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cretich, M" uniqKey="Cretich M">M Cretich</name></author><author><name sortKey="Damin, F" uniqKey="Damin F">F Damin</name></author><author><name sortKey="Pirri, G" uniqKey="Pirri G">G Pirri</name></author><author><name sortKey="Chiari, M" uniqKey="Chiari M">M Chiari</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chandra, H" uniqKey="Chandra H">H Chandra</name></author><author><name sortKey="Srivastava, S" uniqKey="Srivastava S">S Srivastava</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Benjamin, Dc" uniqKey="Benjamin D">DC Benjamin</name></author><author><name sortKey="Perdue, Ss" uniqKey="Perdue S">SS Perdue</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pande, J" uniqKey="Pande J">J Pande</name></author><author><name sortKey="Szewczyk, Mm" uniqKey="Szewczyk M">MM Szewczyk</name></author><author><name sortKey="Grover, Ak" uniqKey="Grover A">AK Grover</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Timmerman, P" uniqKey="Timmerman P">P Timmerman</name></author><author><name sortKey="Puijk, Wc" uniqKey="Puijk W">WC Puijk</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Van Regenmortel, Mh" uniqKey="Van Regenmortel M">MH Van Regenmortel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Geysen, Hm" uniqKey="Geysen H">HM Geysen</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author><author><name sortKey="Barteling, Sj" uniqKey="Barteling S">SJ Barteling</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Houghten, Ra" uniqKey="Houghten R">RA Houghten</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Furka, A" uniqKey="Furka A">A Furka</name></author><author><name sortKey="Sebestyen, F" uniqKey="Sebestyen F">F Sebestyen</name></author><author><name sortKey="Asgedom, M" uniqKey="Asgedom M">M Asgedom</name></author><author><name sortKey="Dibo, G" uniqKey="Dibo G">G Dibo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lam, Ks" uniqKey="Lam K">KS Lam</name></author><author><name sortKey="Salmon, Se" uniqKey="Salmon S">SE Salmon</name></author><author><name sortKey="Hersh, Em" uniqKey="Hersh E">EM Hersh</name></author><author><name sortKey="Hruby, Vj" uniqKey="Hruby V">VJ Hruby</name></author><author><name sortKey="Kazmierski, Wm" uniqKey="Kazmierski W">WM Kazmierski</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fodor, Sp" uniqKey="Fodor S">SP Fodor</name></author><author><name sortKey="Read, Jl" uniqKey="Read J">JL Read</name></author><author><name sortKey="Pirrung, Mc" uniqKey="Pirrung M">MC Pirrung</name></author><author><name sortKey="Stryer, L" uniqKey="Stryer L">L Stryer</name></author><author><name sortKey="Lu, At" uniqKey="Lu A">AT Lu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Frank, R" uniqKey="Frank R">R Frank</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kramer, A" uniqKey="Kramer A">A Kramer</name></author><author><name sortKey="Schneider Mergener, J" uniqKey="Schneider Mergener J">J Schneider-Mergener</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Breitling, F" uniqKey="Breitling F">F Breitling</name></author><author><name sortKey="Felgenhauer, T" uniqKey="Felgenhauer T">T Felgenhauer</name></author><author><name sortKey="Nesterov, A" uniqKey="Nesterov A">A Nesterov</name></author><author><name sortKey="Lindenstruth, V" uniqKey="Lindenstruth V">V Lindenstruth</name></author><author><name sortKey="Stadler, V" uniqKey="Stadler V">V Stadler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Singh Gasson, S" uniqKey="Singh Gasson S">S Singh-Gasson</name></author><author><name sortKey="Green, Rd" uniqKey="Green R">RD Green</name></author><author><name sortKey="Yue, Y" uniqKey="Yue Y">Y Yue</name></author><author><name sortKey="Nelson, C" uniqKey="Nelson C">C Nelson</name></author><author><name sortKey="Blattner, F" uniqKey="Blattner F">F 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sortKey="Forsstrom, B" uniqKey="Forsstrom B">B Forsstrom</name></author><author><name sortKey="Nilsson, P" uniqKey="Nilsson P">P Nilsson</name></author><author><name sortKey="Uhlen, M" uniqKey="Uhlen M">M Uhlen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yxfeldt, G" uniqKey="Yxfeldt G">G Yxfeldt</name></author><author><name sortKey="Froman, G" uniqKey="Froman G">G Froman</name></author><author><name sortKey="Mardh, Pa" uniqKey="Mardh P">PA Mardh</name></author><author><name sortKey="Ward, Me" uniqKey="Ward M">ME Ward</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Timmerman, P" uniqKey="Timmerman P">P Timmerman</name></author><author><name sortKey="Beld, J" uniqKey="Beld J">J Beld</name></author><author><name sortKey="Puijk, Wc" uniqKey="Puijk W">WC Puijk</name></author><author><name sortKey="Meloen, Rh" uniqKey="Meloen R">RH Meloen</name></author></analytic></biblStruct></listBibl></div1></back></TEI></pmc><pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Identification and mapping of linear antibody epitopes in human serum albumin using high-density Peptide arrays.</title><author><name sortKey="Hansen, Lajla Bruntse" sort="Hansen, Lajla Bruntse" uniqKey="Hansen L" first="Lajla Bruntse" last="Hansen">Lajla Bruntse Hansen</name><affiliation wicri:level="3"><nlm:affiliation>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Buus, Soren" sort="Buus, Soren" uniqKey="Buus S" first="Soren" last="Buus">Soren Buus</name></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2013">2013</date><idno type="RBID">pubmed:23894373</idno><idno type="pmid">23894373</idno><idno type="doi">10.1371/journal.pone.0068902</idno><idno type="wicri:Area/PubMed/Corpus">001C40</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001C40</idno><idno type="wicri:Area/PubMed/Curation">001C40</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001C40</idno><idno type="wicri:Area/PubMed/Checkpoint">001B33</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001B33</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Identification and mapping of linear antibody epitopes in human serum albumin using high-density Peptide arrays.</title><author><name sortKey="Hansen, Lajla Bruntse" sort="Hansen, Lajla Bruntse" uniqKey="Hansen L" first="Lajla Bruntse" last="Hansen">Lajla Bruntse Hansen</name><affiliation wicri:level="3"><nlm:affiliation>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen, Denmark.</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>Laboratory of Experimental Immunology, University of Copenhagen, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Buus, Soren" sort="Buus, Soren" uniqKey="Buus S" first="Soren" last="Buus">Soren Buus</name></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name></author></analytic><series><title level="j">PloS one</title><idno type="eISSN">1932-6203</idno><imprint><date when="2013" type="published">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aged</term><term>Aged, 80 and over</term><term>Alzheimer Disease (diagnosis)</term><term>Alzheimer Disease (metabolism)</term><term>Animals</term><term>Brain Mapping</term><term>Case-Control Studies</term><term>Cattle</term><term>Cell Physiological Phenomena</term><term>Cognitive Dysfunction (diagnosis)</term><term>Cognitive Dysfunction (metabolism)</term><term>Female</term><term>Fluorodeoxyglucose F18</term><term>Follow-Up Studies</term><term>Glucose (metabolism)</term><term>Humans</term><term>Longitudinal Studies</term><term>Male</term><term>Middle Aged</term><term>Neural Networks, Computer</term><term>Neuroimaging</term><term>Positron-Emission Tomography</term><term>Rabbits</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adulte d'âge moyen</term><term>Animaux</term><term>Bovins</term><term>Cartographie cérébrale</term><term>Dysfonctionnement cognitif (diagnostic)</term><term>Dysfonctionnement cognitif (métabolisme)</term><term>Femelle</term><term>Fluorodésoxyglucose F18</term><term>Glucose (métabolisme)</term><term>Humains</term><term>Lapins</term><term>Maladie d'Alzheimer (diagnostic)</term><term>Maladie d'Alzheimer (métabolisme)</term><term>Mâle</term><term>Neuroimagerie</term><term>Phénomènes physiologiques cellulaires</term><term>Sujet âgé</term><term>Sujet âgé de 80 ans ou plus</term><term>Tomographie par émission de positons</term><term>Études cas-témoins</term><term>Études de suivi</term><term>Études longitudinales</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glucose</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Fluorodeoxyglucose F18</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Alzheimer Disease</term><term>Cognitive Dysfunction</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Dysfonctionnement cognitif</term><term>Maladie d'Alzheimer</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Alzheimer Disease</term><term>Cognitive Dysfunction</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Dysfonctionnement cognitif</term><term>Glucose</term><term>Maladie d'Alzheimer</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Aged</term><term>Aged, 80 and over</term><term>Animals</term><term>Brain Mapping</term><term>Case-Control Studies</term><term>Cattle</term><term>Cell Physiological Phenomena</term><term>Female</term><term>Follow-Up Studies</term><term>Humans</term><term>Longitudinal Studies</term><term>Male</term><term>Middle Aged</term><term>Neural Networks, Computer</term><term>Neuroimaging</term><term>Positron-Emission Tomography</term><term>Rabbits</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adulte d'âge moyen</term><term>Animaux</term><term>Bovins</term><term>Cartographie cérébrale</term><term>Femelle</term><term>Fluorodésoxyglucose F18</term><term>Humains</term><term>Lapins</term><term>Mâle</term><term>Neuroimagerie</term><term>Phénomènes physiologiques cellulaires</term><term>Sujet âgé</term><term>Sujet âgé de 80 ans ou plus</term><term>Tomographie par émission de positons</term><term>Études cas-témoins</term><term>Études de suivi</term><term>Études longitudinales</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm(2). Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level. </div></front></TEI></pubmed></double></record>Pour manipuler ce document sous Unix (Dilib)
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