A nylon ball solid-phase radioimmunoassay for specific antibodies in human sera. Application to measurement of IgG antibodies to pollen allergens
The principle of the radioallergosorbent test (RAST) has been used to measure IgG antibodies to timothy grass pollen allergens in sera from desensitized allergic subjects. 125I-labeled goat anti-human IgG was used as detector protein. Non-specific binding was eliminated by use of a non-porous nylon...
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| Veröffentlicht in: | Journal of immunological methods 1983-09, Vol.62 (3), p.283-296 |
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| creator | Djurup, R. Søndergaard, I. Minuva, U. Weeke, B. |
| description | The principle of the radioallergosorbent test (RAST) has been used to measure IgG antibodies to timothy grass pollen allergens in sera from desensitized allergic subjects.
125I-labeled goat anti-human IgG was used as detector protein. Non-specific binding was eliminated by use of a non-porous nylon ball an antigen carrier and by use of a special buffer with high ionic strength and pH, containing 1% bovine gamma globulin and 5% normal rabbit serum as ‘balance proteins’. At dilution 1 : 80 non-specific binding was only 0.28% and the binding ratio for a high-titer serum was about 10. By inhibition experiments the assay was demonstrated to be specific for IgG antibodies to timothy grass pollen. The results obtained with this assay correlated statistically significant with those found with double-antibody method (
r
S = 0.68, n = 20,
t = 3.93,
P < 0.001). Serum dilution curves were parallel, indicating that the assay is in allergen excess. The within-assay coefficient of variation ranged from 3.9 to 7.6%; the between-assay coefficient of variation from 8.4 to 19.5%. The assay is very simple to perform, requiring no centrifugation. The allergen-coated balls are stable for at least 3 months. The assay should be applicable to measurement of IgG antibodies and IgG subclass antibodies to any protein antigen of interest. |
| doi_str_mv | 10.1016/0022-1759(83)90172-2 |
| format | Article |
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125I-labeled goat anti-human IgG was used as detector protein. Non-specific binding was eliminated by use of a non-porous nylon ball an antigen carrier and by use of a special buffer with high ionic strength and pH, containing 1% bovine gamma globulin and 5% normal rabbit serum as ‘balance proteins’. At dilution 1 : 80 non-specific binding was only 0.28% and the binding ratio for a high-titer serum was about 10. By inhibition experiments the assay was demonstrated to be specific for IgG antibodies to timothy grass pollen. The results obtained with this assay correlated statistically significant with those found with double-antibody method (
r
S = 0.68, n = 20,
t = 3.93,
P < 0.001). Serum dilution curves were parallel, indicating that the assay is in allergen excess. The within-assay coefficient of variation ranged from 3.9 to 7.6%; the between-assay coefficient of variation from 8.4 to 19.5%. The assay is very simple to perform, requiring no centrifugation. The allergen-coated balls are stable for at least 3 months. The assay should be applicable to measurement of IgG antibodies and IgG subclass antibodies to any protein antigen of interest.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/0022-1759(83)90172-2</identifier><identifier>PMID: 6604105</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Antibodies - analysis ; Antibodies, Anti-Idiotypic - immunology ; Binding Sites, Antibody ; Biological and medical sciences ; Buffers ; Cross Reactions ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Humans ; Hydrogen-Ion Concentration ; IgG antibody to pollen allergen ; IgG RAST ; Immunoglobulin G - analysis ; immunotherapy ; Kinetics ; Microspheres ; Molecular immunology ; Osmolar Concentration ; Pollen - immunology ; Protein Binding ; Radioimmunoassay - methods ; Radioimmunoassay - standards ; solid-phase radioimmunoassay ; Techniques ; timothy grass pollen</subject><ispartof>Journal of immunological methods, 1983-09, Vol.62 (3), p.283-296</ispartof><rights>1983</rights><rights>1984 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-6b24233664e29629b636896a4ac1a056262a59b67cd02f34b7de08421e347c2d3</citedby><cites>FETCH-LOGICAL-c386t-6b24233664e29629b636896a4ac1a056262a59b67cd02f34b7de08421e347c2d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0022-1759(83)90172-2$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9342848$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6604105$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Djurup, R.</creatorcontrib><creatorcontrib>Søndergaard, I.</creatorcontrib><creatorcontrib>Minuva, U.</creatorcontrib><creatorcontrib>Weeke, B.</creatorcontrib><title>A nylon ball solid-phase radioimmunoassay for specific antibodies in human sera. Application to measurement of IgG antibodies to pollen allergens</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>The principle of the radioallergosorbent test (RAST) has been used to measure IgG antibodies to timothy grass pollen allergens in sera from desensitized allergic subjects.
125I-labeled goat anti-human IgG was used as detector protein. Non-specific binding was eliminated by use of a non-porous nylon ball an antigen carrier and by use of a special buffer with high ionic strength and pH, containing 1% bovine gamma globulin and 5% normal rabbit serum as ‘balance proteins’. At dilution 1 : 80 non-specific binding was only 0.28% and the binding ratio for a high-titer serum was about 10. By inhibition experiments the assay was demonstrated to be specific for IgG antibodies to timothy grass pollen. The results obtained with this assay correlated statistically significant with those found with double-antibody method (
r
S = 0.68, n = 20,
t = 3.93,
P < 0.001). Serum dilution curves were parallel, indicating that the assay is in allergen excess. The within-assay coefficient of variation ranged from 3.9 to 7.6%; the between-assay coefficient of variation from 8.4 to 19.5%. The assay is very simple to perform, requiring no centrifugation. The allergen-coated balls are stable for at least 3 months. The assay should be applicable to measurement of IgG antibodies and IgG subclass antibodies to any protein antigen of interest.</description><subject>Antibodies - analysis</subject><subject>Antibodies, Anti-Idiotypic - immunology</subject><subject>Binding Sites, Antibody</subject><subject>Biological and medical sciences</subject><subject>Buffers</subject><subject>Cross Reactions</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>IgG antibody to pollen allergen</subject><subject>IgG RAST</subject><subject>Immunoglobulin G - analysis</subject><subject>immunotherapy</subject><subject>Kinetics</subject><subject>Microspheres</subject><subject>Molecular immunology</subject><subject>Osmolar Concentration</subject><subject>Pollen - immunology</subject><subject>Protein Binding</subject><subject>Radioimmunoassay - methods</subject><subject>Radioimmunoassay - standards</subject><subject>solid-phase radioimmunoassay</subject><subject>Techniques</subject><subject>timothy grass pollen</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1983</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc2KFDEUhYMoYzv6BgpZiOiixvxUp6o2A82g48CAG12HW6lbM5FUUuZWCf0YvrFpu2lcuUrI_c454VzGXktxJYU0H4VQqpLNtnvf6g-dkI2q1BO2kW25NJ3YPmWbM_KcvSD6IYSQwogLdmGMqKXYbtjvHY_7kCLvIQROKfihmh-BkGcYfPLTtMYERLDnY8qcZnR-9I5DXHyfBo_EfeSP6wSRE2a44rt5Dt7B4ovpkviEQGvGCePC08jvHm7_1RZgTiFg5CUe8wNGesmejRAIX53OS_b986dvN1-q-6-3dze7-8rp1iyV6VWttDamRtUZ1fVGm7YzUIOTILZGGQXb8tq4QahR130zoGhrJVHXjVODvmTvjr5zTj9XpMVOnhyGABHTSrYVRrV11xawPoIuJ6KMo52znyDvrRT2sAl7qNkearattn83YVWRvTn5r_2Ew1l0qr7M357mQA7CmCE6T2es03WJP6RfHzEsXfzymC05j9Hh4DO6xQ7J__8ffwD826W0</recordid><startdate>19830916</startdate><enddate>19830916</enddate><creator>Djurup, R.</creator><creator>Søndergaard, I.</creator><creator>Minuva, U.</creator><creator>Weeke, B.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19830916</creationdate><title>A nylon ball solid-phase radioimmunoassay for specific antibodies in human sera. Application to measurement of IgG antibodies to pollen allergens</title><author>Djurup, R. ; Søndergaard, I. ; Minuva, U. ; Weeke, B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-6b24233664e29629b636896a4ac1a056262a59b67cd02f34b7de08421e347c2d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1983</creationdate><topic>Antibodies - analysis</topic><topic>Antibodies, Anti-Idiotypic - immunology</topic><topic>Binding Sites, Antibody</topic><topic>Biological and medical sciences</topic><topic>Buffers</topic><topic>Cross Reactions</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>IgG antibody to pollen allergen</topic><topic>IgG RAST</topic><topic>Immunoglobulin G - analysis</topic><topic>immunotherapy</topic><topic>Kinetics</topic><topic>Microspheres</topic><topic>Molecular immunology</topic><topic>Osmolar Concentration</topic><topic>Pollen - immunology</topic><topic>Protein Binding</topic><topic>Radioimmunoassay - methods</topic><topic>Radioimmunoassay - standards</topic><topic>solid-phase radioimmunoassay</topic><topic>Techniques</topic><topic>timothy grass pollen</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Djurup, R.</creatorcontrib><creatorcontrib>Søndergaard, I.</creatorcontrib><creatorcontrib>Minuva, U.</creatorcontrib><creatorcontrib>Weeke, B.</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>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Djurup, R.</au><au>Søndergaard, I.</au><au>Minuva, U.</au><au>Weeke, B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A nylon ball solid-phase radioimmunoassay for specific antibodies in human sera. Application to measurement of IgG antibodies to pollen allergens</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1983-09-16</date><risdate>1983</risdate><volume>62</volume><issue>3</issue><spage>283</spage><epage>296</epage><pages>283-296</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>The principle of the radioallergosorbent test (RAST) has been used to measure IgG antibodies to timothy grass pollen allergens in sera from desensitized allergic subjects.
125I-labeled goat anti-human IgG was used as detector protein. Non-specific binding was eliminated by use of a non-porous nylon ball an antigen carrier and by use of a special buffer with high ionic strength and pH, containing 1% bovine gamma globulin and 5% normal rabbit serum as ‘balance proteins’. At dilution 1 : 80 non-specific binding was only 0.28% and the binding ratio for a high-titer serum was about 10. By inhibition experiments the assay was demonstrated to be specific for IgG antibodies to timothy grass pollen. The results obtained with this assay correlated statistically significant with those found with double-antibody method (
r
S = 0.68, n = 20,
t = 3.93,
P < 0.001). Serum dilution curves were parallel, indicating that the assay is in allergen excess. The within-assay coefficient of variation ranged from 3.9 to 7.6%; the between-assay coefficient of variation from 8.4 to 19.5%. The assay is very simple to perform, requiring no centrifugation. The allergen-coated balls are stable for at least 3 months. The assay should be applicable to measurement of IgG antibodies and IgG subclass antibodies to any protein antigen of interest.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>6604105</pmid><doi>10.1016/0022-1759(83)90172-2</doi><tpages>14</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-1759 |
| ispartof | Journal of immunological methods, 1983-09, Vol.62 (3), p.283-296 |
| issn | 0022-1759 1872-7905 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_80628498 |
| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Antibodies - analysis Antibodies, Anti-Idiotypic - immunology Binding Sites, Antibody Biological and medical sciences Buffers Cross Reactions Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Hydrogen-Ion Concentration IgG antibody to pollen allergen IgG RAST Immunoglobulin G - analysis immunotherapy Kinetics Microspheres Molecular immunology Osmolar Concentration Pollen - immunology Protein Binding Radioimmunoassay - methods Radioimmunoassay - standards solid-phase radioimmunoassay Techniques timothy grass pollen |
| title | A nylon ball solid-phase radioimmunoassay for specific antibodies in human sera. Application to measurement of IgG antibodies to pollen allergens |
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