Abstract:
:We describe the preparation of a new rat monoclonal antibody (CRC1) to the N-terminal sequence of the 43 kDa subunit of human ovarian inhibin, and its use together with other anti-peptide monoclonal antibodies, in two-site immunoassays for the detection of inhibin-related material in biological fluids. The Fab fraction of a mouse monoclonal antibody (R1) to the N-terminal portion of the 20 kDa alpha subunit, coupled to alkaline phosphatase, was used for detection, and either CRC1 or a monoclonal antibody (E4) to the beta-A subunit were used as capture antibodies. The E4/R1 combination, expected to measure dimeric bioactive inhibin, could detect less than 2 pg/ml of recombinant inhibin in diluent, gave good recovery of activity spiked into human blood, and could measure significant levels of immunoreactivity in sera from women undergoing ovulation induction, and in some normal women. Sera from post-menopausal women contained undetectable levels. Apparent inhibin levels in human follicular fluid were increased six-fold by pretreatment with 8 M urea, suggesting masking of epitopes in this fluid. Activin cross-reactivity in the assay was 0.05%. The R1/CRC1 assay, expected to measure only large molecular weight forms of inhibin or its alpha subunit, could detect immunoreactivity in human FF diluted 50,000-fold, and in all sera tested, although the levels in the hyperovulated women were higher. By contrast to the E4/R1 assay much of the immunoreactivity was labile during the clotting process, or subsequent assay, and reliable measurements on blood with this assay will require special sample collection procedures. These results demonstrate the value of anti-peptide monoclonal antibodies in the study of inhibin, and the results obtained with CRC1 show that antibodies useful for immunoassays can sometimes be obtained without the purified target molecule being available for immunization or screening.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Groome N,O'Brien Mdoi
10.1016/0022-1759(93)90342-5subject
Has Abstractpub_date
1993-10-15 00:00:00pages
167-76issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(93)90342-5journal_volume
165pub_type
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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更新日期:1997-01-15 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1993-01-14 00:00:00
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1985-11-28 00:00:00
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journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1987-12-04 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1980-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:2016-08-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90042-3
更新日期:1980-01-01 00:00:00
abstract::A solid-phase enzyme immunoassay for the measurement of anti-acetylcholine receptor antibodies in the sera of patients with myasthenia gravis is reported. Sufficient amounts of acetylcholine receptor for the sensitive detection of anti-acetylcholine receptor antibody were directly fixed to Costar serocluster 96-well E...
journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1984-10-26 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(81)90089-2
更新日期:1981-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:1990-08-07 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
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更新日期:1996-09-13 00:00:00