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קהילה:
אסיף מאגר המחקר החקלאי
פותח על ידי קלירמאש פתרונות בע"מ -
Monitoring of progestins: Development of immunochemical methods for purification and detection of levonorgestrel
Year:
2010
Source of publication :
Analytica Chimica Acta
Authors :
אלטשטיין, מרים
;
.
ברדוגו, מיכל
;
.
ברונשטיין, אליסה
;
.
שלו, מורן
;
.
Volume :
665
Co-Authors:
Shalev, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Bardugo, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Nudelman, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Krol, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Schlesinger, H., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Bronshtein, A., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Altstein, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Facilitators :
From page:
176
To page:
184
(
Total pages:
9
)
Abstract:
A polyclonal antibody (Ab) for the progestin levonorgestrel (LNG) was generated, and immunochemical assays for its detection, clean-up and concentration were developed. A highly specific microplate diagnostic assay for the detection of LNG was developed that used the enzyme linked immunosorbent assay (ELISA) method. The LNG ELISA developed was sensitive and reproducible; it exhibited I50 and I20 values of 3.3±1.8ngmL-1 and 0.6±0.4ngmL-1, respectively, and the Abs did not cross react with any of the tested steroid hormones. The above Abs were used to develop a sol-gel-based immunoaffinity purification (IAP) method for concentration and clean-up of LNG that is compatible with subsequent immunochemical or instrumental chemical analytical procedures, such as liquid chromatography followed by mass spectrometry (LC-MS/MS). Development of the columns included successful entrapment of Abs within a tetramethoxysilane (TMOS)-based SiO2 polymer network. The Abs could bind the free analyte from solution, and the bound analyte could be easily eluted from the sol-gel matrix at high recoveries. The Ab selectivity towards the antigen was high, in both ELISA and the sol-gel columns, but the entrapped Abs cross-reacted with two other steroid hormones - ethynylestradiol (EE2) and nortestosterone (NT) - which share similar epitopes with LNG, despite the lack of cross reactivity in the ELISA. The validity of the method was investigated by LC-MS/MS and a good analytical correlation was obtained. © 2010 Elsevier B.V.
Note:
Related Files :
Chromatography, High Pressure Liquid
Endocrinology
enzymes
gestagen
hormones
nandrolone
Residue monitoring
עוד תגיות
תוכן קשור
More details
DOI :
10.1016/j.aca.2010.03.029
Article number:
0
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
21064
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:41
Scientific Publication
Monitoring of progestins: Development of immunochemical methods for purification and detection of levonorgestrel
665
Shalev, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Bardugo, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Nudelman, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Krol, A., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Schlesinger, H., Analyst Research Laboratories, Rabin Park, Hamada St. 12, Rehovot, 76703, Israel
Bronshtein, A., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Altstein, M., Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan 50250, Israel
Monitoring of progestins: Development of immunochemical methods for purification and detection of levonorgestrel
A polyclonal antibody (Ab) for the progestin levonorgestrel (LNG) was generated, and immunochemical assays for its detection, clean-up and concentration were developed. A highly specific microplate diagnostic assay for the detection of LNG was developed that used the enzyme linked immunosorbent assay (ELISA) method. The LNG ELISA developed was sensitive and reproducible; it exhibited I50 and I20 values of 3.3±1.8ngmL-1 and 0.6±0.4ngmL-1, respectively, and the Abs did not cross react with any of the tested steroid hormones. The above Abs were used to develop a sol-gel-based immunoaffinity purification (IAP) method for concentration and clean-up of LNG that is compatible with subsequent immunochemical or instrumental chemical analytical procedures, such as liquid chromatography followed by mass spectrometry (LC-MS/MS). Development of the columns included successful entrapment of Abs within a tetramethoxysilane (TMOS)-based SiO2 polymer network. The Abs could bind the free analyte from solution, and the bound analyte could be easily eluted from the sol-gel matrix at high recoveries. The Ab selectivity towards the antigen was high, in both ELISA and the sol-gel columns, but the entrapped Abs cross-reacted with two other steroid hormones - ethynylestradiol (EE2) and nortestosterone (NT) - which share similar epitopes with LNG, despite the lack of cross reactivity in the ELISA. The validity of the method was investigated by LC-MS/MS and a good analytical correlation was obtained. © 2010 Elsevier B.V.
Scientific Publication
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