A sensitive two-site enzyme-linked immunosorbent assay for measurement of the major Alternaria alternata allergen Alt a 1
Alt a 1 is the major allergen in Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality. To develop an Alt a 1-speci...
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| Veröffentlicht in: | Annals of allergy, asthma, & immunology asthma, & immunology, 2003-05, Vol.90 (5), p.529-535 |
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| container_title | Annals of allergy, asthma, & immunology |
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| creator | Asturias, Juan A. Arilla, M. Carmen Ibarrola, Ignacio Eraso, Elena González-Rioja, Roberto Martínez, Alberto |
| description | Alt a 1 is the major allergen in
Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality.
To develop an Alt a 1-specific assay and to evaluate the correlation of Alt a 1 content with the IgE-binding activity of
A. alternata extracts.
Recombinant Alt a 1 was produced as nonfusion protein from a polymerase chain reaction-cloned complementary DNA Alt a 1 sequence. Natural Alt a 1 was purified from spent culture medium. Monoclonal and polyclonal antibodies directed to Alt a 1 were produced and used to construct a specific Alt a 1 enzyme-linked immunosorbent assay (ELISA).
The ELISA developed was highly reproducible and sensitive, with a detection limit lower than 0.5 ng/mL and a practical working range of 0.5 to 50 ng/mL. The assay was able to detect an Alt a 1-like protein in
Stemphylium extracts. Identical parallel dose-response curves were observed when natural Alt a 1 and recombinant Alt a 1 were used as standard. A good correlation was obtained between Alt a 1 content of 13
A. alternata extracts and their IgE-binding activity. Alt a 1 was responsible for 70% of the IgE-binding activity of the whole extract.
This sensitive and specific Alt a 1 assay allows the quantification of this major mold allergen and represents a useful tool for the standardization of
A. alternata extracts in mass units. It also provides a reliable indication of the allergenic activity of the whole extract. |
| doi_str_mv | 10.1016/S1081-1206(10)61846-7 |
| format | Article |
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Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality.
To develop an Alt a 1-specific assay and to evaluate the correlation of Alt a 1 content with the IgE-binding activity of
A. alternata extracts.
Recombinant Alt a 1 was produced as nonfusion protein from a polymerase chain reaction-cloned complementary DNA Alt a 1 sequence. Natural Alt a 1 was purified from spent culture medium. Monoclonal and polyclonal antibodies directed to Alt a 1 were produced and used to construct a specific Alt a 1 enzyme-linked immunosorbent assay (ELISA).
The ELISA developed was highly reproducible and sensitive, with a detection limit lower than 0.5 ng/mL and a practical working range of 0.5 to 50 ng/mL. The assay was able to detect an Alt a 1-like protein in
Stemphylium extracts. Identical parallel dose-response curves were observed when natural Alt a 1 and recombinant Alt a 1 were used as standard. A good correlation was obtained between Alt a 1 content of 13
A. alternata extracts and their IgE-binding activity. Alt a 1 was responsible for 70% of the IgE-binding activity of the whole extract.
This sensitive and specific Alt a 1 assay allows the quantification of this major mold allergen and represents a useful tool for the standardization of
A. alternata extracts in mass units. It also provides a reliable indication of the allergenic activity of the whole extract.</description><identifier>ISSN: 1081-1206</identifier><identifier>EISSN: 1534-4436</identifier><identifier>DOI: 10.1016/S1081-1206(10)61846-7</identifier><identifier>PMID: 12775134</identifier><identifier>CODEN: ANAEA3</identifier><language>eng</language><publisher>McLean, VA: Elsevier Inc</publisher><subject>Allergens - analysis ; Allergens - genetics ; Allergens - immunology ; Allergological tests ; Alternaria - genetics ; Alternaria - immunology ; Alternaria alternata ; Amino Acid Sequence ; Animals ; Antibodies, Fungal - biosynthesis ; Antibodies, Monoclonal - immunology ; Antigens, Plant ; Base Sequence ; Biological and medical sciences ; Dose-Response Relationship, Immunologic ; Enzyme-Linked Immunosorbent Assay - methods ; Female ; Fungal Proteins - analysis ; Fungal Proteins - genetics ; Fungal Proteins - immunology ; Humans ; Immunoblotting ; Immunoglobulin E - immunology ; Immunological methods for diagnosis and exploration ; Immunopathology ; Medical sciences ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Rabbits ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Fungal - chemistry ; RNA, Fungal - genetics ; Sensitivity and Specificity</subject><ispartof>Annals of allergy, asthma, & immunology, 2003-05, Vol.90 (5), p.529-535</ispartof><rights>2003 American College of Allergy, Asthma & Immunology</rights><rights>2003 INIST-CNRS</rights><rights>Copyright American College of Allergy and Immunology May 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c449t-98e61b236292e2656d6c0d5f366f57b65d430ba1213648f6e8510badffb735f83</citedby><cites>FETCH-LOGICAL-c449t-98e61b236292e2656d6c0d5f366f57b65d430ba1213648f6e8510badffb735f83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S1081-1206(10)61846-7$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14852844$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12775134$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Asturias, Juan A.</creatorcontrib><creatorcontrib>Arilla, M. Carmen</creatorcontrib><creatorcontrib>Ibarrola, Ignacio</creatorcontrib><creatorcontrib>Eraso, Elena</creatorcontrib><creatorcontrib>González-Rioja, Roberto</creatorcontrib><creatorcontrib>Martínez, Alberto</creatorcontrib><title>A sensitive two-site enzyme-linked immunosorbent assay for measurement of the major Alternaria alternata allergen Alt a 1</title><title>Annals of allergy, asthma, & immunology</title><addtitle>Ann Allergy Asthma Immunol</addtitle><description>Alt a 1 is the major allergen in
Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality.
To develop an Alt a 1-specific assay and to evaluate the correlation of Alt a 1 content with the IgE-binding activity of
A. alternata extracts.
Recombinant Alt a 1 was produced as nonfusion protein from a polymerase chain reaction-cloned complementary DNA Alt a 1 sequence. Natural Alt a 1 was purified from spent culture medium. Monoclonal and polyclonal antibodies directed to Alt a 1 were produced and used to construct a specific Alt a 1 enzyme-linked immunosorbent assay (ELISA).
The ELISA developed was highly reproducible and sensitive, with a detection limit lower than 0.5 ng/mL and a practical working range of 0.5 to 50 ng/mL. The assay was able to detect an Alt a 1-like protein in
Stemphylium extracts. Identical parallel dose-response curves were observed when natural Alt a 1 and recombinant Alt a 1 were used as standard. A good correlation was obtained between Alt a 1 content of 13
A. alternata extracts and their IgE-binding activity. Alt a 1 was responsible for 70% of the IgE-binding activity of the whole extract.
This sensitive and specific Alt a 1 assay allows the quantification of this major mold allergen and represents a useful tool for the standardization of
A. alternata extracts in mass units. It also provides a reliable indication of the allergenic activity of the whole extract.</description><subject>Allergens - analysis</subject><subject>Allergens - genetics</subject><subject>Allergens - immunology</subject><subject>Allergological tests</subject><subject>Alternaria - genetics</subject><subject>Alternaria - immunology</subject><subject>Alternaria alternata</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibodies, Fungal - biosynthesis</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigens, Plant</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Dose-Response Relationship, Immunologic</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Female</subject><subject>Fungal Proteins - analysis</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - immunology</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Immunoglobulin E - immunology</subject><subject>Immunological methods for diagnosis and exploration</subject><subject>Immunopathology</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Molecular Sequence Data</subject><subject>Rabbits</subject><subject>Reproducibility of Results</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Fungal - chemistry</subject><subject>RNA, Fungal - genetics</subject><subject>Sensitivity and Specificity</subject><issn>1081-1206</issn><issn>1534-4436</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhi0Eou3CTwBZSCA4BDz-inOqVlX5kCpxAM6Rk4zBS2IXOylafj1Od1ElLj359cwzo5l5CXkG7C0w0O--ADNQAWf6NbA3GozUVf2AnIISspJS6IdF_0NOyFnOO8YYGC0ekxPgda1AyFOy39KMIfvZ3yCdf8eqSKQY_uwnrEYffuJA_TQtIeaYOgwztTnbPXUx0QltXhJOazQ6Ov9AOtldSWzHGVOwyVtqD3Je1YjpO4Y1Sy2FJ-SRs2PGp8d3Q769v_x68bG6-vzh08X2quqlbOaqMaih40LzhiPXSg-6Z4NyQmun6k6rQQrWWeAgtDROo1FQ_oNzXS2UM2JDXh36Xqf4a8E8t5PPPY6jDRiX3NZCKOCC3wuCaaRpGlbAF_-Bu7iUJcfccsZro1W58oaoA9SnmHNC114nP9m0b4G1q4PtrYPtas8aunWwDLMhz4_Nl27C4a7qaFkBXh4Bm3s7umRD7_MdJ43iRq7c-YHDctwbj6nNvcfQ4-AT9nM7RH_PKH8Bqau3mQ</recordid><startdate>20030501</startdate><enddate>20030501</enddate><creator>Asturias, Juan A.</creator><creator>Arilla, M. 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Carmen ; Ibarrola, Ignacio ; Eraso, Elena ; González-Rioja, Roberto ; Martínez, Alberto</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c449t-98e61b236292e2656d6c0d5f366f57b65d430ba1213648f6e8510badffb735f83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Allergens - analysis</topic><topic>Allergens - genetics</topic><topic>Allergens - immunology</topic><topic>Allergological tests</topic><topic>Alternaria - genetics</topic><topic>Alternaria - immunology</topic><topic>Alternaria alternata</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibodies, Fungal - biosynthesis</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antigens, Plant</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Dose-Response Relationship, Immunologic</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Female</topic><topic>Fungal Proteins - analysis</topic><topic>Fungal Proteins - genetics</topic><topic>Fungal Proteins - immunology</topic><topic>Humans</topic><topic>Immunoblotting</topic><topic>Immunoglobulin E - immunology</topic><topic>Immunological methods for diagnosis and exploration</topic><topic>Immunopathology</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Molecular Sequence Data</topic><topic>Rabbits</topic><topic>Reproducibility of Results</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Fungal - chemistry</topic><topic>RNA, Fungal - genetics</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Asturias, Juan A.</creatorcontrib><creatorcontrib>Arilla, M. Carmen</creatorcontrib><creatorcontrib>Ibarrola, Ignacio</creatorcontrib><creatorcontrib>Eraso, Elena</creatorcontrib><creatorcontrib>González-Rioja, Roberto</creatorcontrib><creatorcontrib>Martínez, Alberto</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Premium</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Annals of allergy, asthma, & immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Asturias, Juan A.</au><au>Arilla, M. Carmen</au><au>Ibarrola, Ignacio</au><au>Eraso, Elena</au><au>González-Rioja, Roberto</au><au>Martínez, Alberto</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A sensitive two-site enzyme-linked immunosorbent assay for measurement of the major Alternaria alternata allergen Alt a 1</atitle><jtitle>Annals of allergy, asthma, & immunology</jtitle><addtitle>Ann Allergy Asthma Immunol</addtitle><date>2003-05-01</date><risdate>2003</risdate><volume>90</volume><issue>5</issue><spage>529</spage><epage>535</epage><pages>529-535</pages><issn>1081-1206</issn><eissn>1534-4436</eissn><coden>ANAEA3</coden><abstract>Alt a 1 is the major allergen in
Alternaria alternata, one of the most important fungi associated with allergic diseases. Mold allergenic extracts show considerable heterogeneity, and thus accurate standardization of these extracts is essential to guarantee their quality.
To develop an Alt a 1-specific assay and to evaluate the correlation of Alt a 1 content with the IgE-binding activity of
A. alternata extracts.
Recombinant Alt a 1 was produced as nonfusion protein from a polymerase chain reaction-cloned complementary DNA Alt a 1 sequence. Natural Alt a 1 was purified from spent culture medium. Monoclonal and polyclonal antibodies directed to Alt a 1 were produced and used to construct a specific Alt a 1 enzyme-linked immunosorbent assay (ELISA).
The ELISA developed was highly reproducible and sensitive, with a detection limit lower than 0.5 ng/mL and a practical working range of 0.5 to 50 ng/mL. The assay was able to detect an Alt a 1-like protein in
Stemphylium extracts. Identical parallel dose-response curves were observed when natural Alt a 1 and recombinant Alt a 1 were used as standard. A good correlation was obtained between Alt a 1 content of 13
A. alternata extracts and their IgE-binding activity. Alt a 1 was responsible for 70% of the IgE-binding activity of the whole extract.
This sensitive and specific Alt a 1 assay allows the quantification of this major mold allergen and represents a useful tool for the standardization of
A. alternata extracts in mass units. It also provides a reliable indication of the allergenic activity of the whole extract.</abstract><cop>McLean, VA</cop><pub>Elsevier Inc</pub><pmid>12775134</pmid><doi>10.1016/S1081-1206(10)61846-7</doi><tpages>7</tpages></addata></record> |
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| ispartof | Annals of allergy, asthma, & immunology, 2003-05, Vol.90 (5), p.529-535 |
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| subjects | Allergens - analysis Allergens - genetics Allergens - immunology Allergological tests Alternaria - genetics Alternaria - immunology Alternaria alternata Amino Acid Sequence Animals Antibodies, Fungal - biosynthesis Antibodies, Monoclonal - immunology Antigens, Plant Base Sequence Biological and medical sciences Dose-Response Relationship, Immunologic Enzyme-Linked Immunosorbent Assay - methods Female Fungal Proteins - analysis Fungal Proteins - genetics Fungal Proteins - immunology Humans Immunoblotting Immunoglobulin E - immunology Immunological methods for diagnosis and exploration Immunopathology Medical sciences Mice Mice, Inbred BALB C Molecular Sequence Data Rabbits Reproducibility of Results Reverse Transcriptase Polymerase Chain Reaction RNA, Fungal - chemistry RNA, Fungal - genetics Sensitivity and Specificity |
| title | A sensitive two-site enzyme-linked immunosorbent assay for measurement of the major Alternaria alternata allergen Alt a 1 |
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