Chemiluminescence immunoassay of plasma progesterone, with progesterone-acridinium ester used as the labeled antigen.

1985 ◽  
Vol 31 (10) ◽  
pp. 1664-1668 ◽  
Author(s):  
A P Richardson ◽  
J B Kim ◽  
G J Barnard ◽  
W P Collins ◽  
F McCapra
Keyword(s):  
Solid Phase ◽  
Light Yield ◽  
Progesterone Concentration ◽  
Chemiluminescence Immunoassay ◽  
Plasma Progesterone ◽  
Binding Reaction ◽  
Dilute Sodium Hydroxide ◽  
Plasma Extract ◽  
Acridinium Ester ◽  
Venous Plasma

Abstract This simple solid-phase chemiluminescence immunoassay for measurement of progesterone in extracts of venous plasma has sensitivity and precision similar to that of conventional radioimmunoassay with use of a tritiated antigen. The labeled antigen, 11 alpha-progesteryl-2-succinoyltyramine-4-(10-methyl)-acridini um-9-carboxylate, and a monoclonal antibody to progesterone-11 alpha-succinyl-bovine serum albumin are incubated with a 100-microL aliquot of plasma extract (equivalent to 20 microL of plasma) and 50 microL of a suspension of an IgG fraction of a donkey antiserum to mouse immunoglobulins, covalently attached to cellulose particles. After the antibody-binding reaction (60 min at 4 degrees C), 1 mL of phosphate buffer is added to each tube, the tubes are centrifuged (5 min, 1500 X g), and the supernatant fluid is aspirated. The washing step is repeated and diluted hydrochloric acid (50 mmol/L, 50 microL) is added to the pellet. Luminescence is initiated by oxidation with dilute sodium hydroxide/hydrogen peroxide. The signal is integrated over 10 s. The light yield is inversely proportional to the progesterone concentration in the standard or sample.

Measurement of plasma estradiol-17 beta by solid-phase chemiluminescence immunoassay.

1982 ◽  
Vol 28 (5) ◽  
pp. 1120-1124 ◽  
Author(s):  
J B Kim ◽  
G J Barnard ◽  
W P Collins ◽  
F Kohen ◽  
H R Lindner ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Light Yield ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
Plasma Estradiol ◽  
Precision And Accuracy ◽  
Sensitivity Specificity ◽  
Venous Plasma

Abstract We describe a simple, solid-phase chemiluminescence immunoassay for the measurement of estradiol-17 beta in extracts of peripheral venous plasma. The method has similar sensitivity, specificity, precision, and accuracy to a conventional radioimmunoassay with a tritiated antigen. An immunoglobulin G fraction od monoclonal antibodies to estradiol-6-carboyxmethyl oxime-bovine serum albumin is passively adsorbed onto the walls of polypropylene tubes. The labeled antigen is estradiol-6-carboxymethyl oxime-aminobutylethyl isoluminol. After the binding reaction (1 h at 22 degrees C), the solution is removed by aspiration (400 microliters). Sodium hydroxide (5 mol/L, 300 microliters) is added and the mixture incubated for 30 min at 37 degrees C. Luminescence is initiated by oxidation of the label with microperoxidase/hydrogen peroxide and the signal integrated for 10 s. The light yield is inversely proportional to the concentration of estradiol in the standard or sample.

Measurement of Serum Thyroxine by Solid-Phase Chemiluminescence Immunoassay

1983 ◽  
Vol 20 (2) ◽  
pp. 100-104 ◽  
Author(s):  
D A Weerasekera ◽  
J B Kim ◽  
G J Barnard ◽  
W P Collins
Keyword(s):  
Hydrogen Peroxide ◽  
Sodium Hydroxide ◽  
Solid Phase ◽  
Light Yield ◽  
Antibody Binding ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
Serum Thyroxine ◽  
Total Thyroxine

Descriptions are given of two solid-phase chemiluminescence immunoassays for the measurement of total thyroxine in serum. The antibodies were either attached to small uniform plastic microspheres (method 1) or passively adsorbed to antibody-coated tubes (method 2). The labelled antigen was thyroxine-aminobutyl ethyl isoluminol. After the antibody-binding reaction the antibody-bound fraction was washed, sodium hydroxide was added, and the mixture was incubated. Luminescence was initiated by oxidation of the label with microperoxidase/hydrogen peroxide and the signal integrated for 10 seconds. The light yield is inversely proportional to the concentration of thyroxine in the standard or sample. Both methods have similar sensitivity and precision to that obtained by a conventional radioimmunoassay.

Chemiluminescence Immunoassay for Progesterone in Plasma Incorporating Acridinium Ester Labelled Antigen

1988 ◽  
Vol 25 (1) ◽  
pp. 27-34 ◽  
Author(s):  
S A Miller ◽  
M S Morton ◽  
A Turkes
Keyword(s):  
Mass Spectrometry ◽  
Cyanogen Bromide ◽  
Solid Phase ◽  
Gas Chromatography Mass Spectrometry ◽  
Chemiluminescence Immunoassay ◽  
Validation Criteria ◽  
Bovine Serum Albumin Conjugate ◽  
Assay Tube ◽  
Acridinium Ester ◽  
Good Agreement

A sensitive, solid-phase chemiluminescence immunoassay suitable for determining progesterone concentrations in plasma has been developed. The solid-phase antiserum was prepared by coupling a monoclonal progesterone-antibody, raised against a progesterone-11α-hemisuccinyl/bovine serum albumin conjugate, to cyanogen bromide activated cellulose. An 11α-progesteryl-2-carboxymethyltyramine-4-(10-methyl)-acridinium-9-carboxylate conjugate was used as the chemiluminescent label. The assay had a lower limit of sensitivity of 3 pg/assay tube and satisfied accepted validation criteria. Progesterone concentrations determined by chemiluminescence assay were in good agreement not only with a radioimmunoassay in routine use but also with a gas chromatography-mass spectrometry procedure.

Monitoring ovarian function by a solid-phase chemiluminescence immunoassay

1982 ◽  
Vol 101 (2) ◽  
pp. 254-263 ◽  
Author(s):  
D. A. Weerasekera ◽  
J. B. Kim ◽  
G. J. Barnard ◽  
W. P. Collins ◽  
F. Kohen ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Ovarian Function ◽  
Solid Phase ◽  
Early Morning ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
Menstrual Cycles ◽  
The Mean ◽  
Assay Precision ◽  
Good Agreement

Abstract. Ovarian function in women may be monitored by a simple, solid-phase, chemiluminescence immunoassay for the measurement of oestrone-3-glucuronide in diluted urine. An IgG fraction of antiserum to oestrone-3-glucuronyl-6-bovine serum albumin is passively adsorbed to the walls of polystyrene tubes. Oestrone-3-glucuronyl-6-aminoethyl-ethyl-isoluminol is the labelled antigen. Daily samples of early morning urine are diluted in buffer (1:100; v/v), and 100 μl removed, in duplicate, for assay. After the binding reaction (1 h at 4°C), the solution is removed by aspiration. The antibody-bound fraction is washed once with buffer (400 μl). Sodium hydroxide (2n, 200 μl) is added and the mixture incubated for 60 min at 22°C. Luminescence is initiated by oxidation of the label with microperoxidase/hydrogen peroxide and the signal integrated for 10s. An evaluation of the method gave the following values: sensitivity of calibration curve 3.12 ± 0.75 pg/tube (mean ± sd). The intra-assay precision (CV%) was 9.52 (20 replicates; 38.4 ± 3.66 nmol/l) and 8.61 (15 replicates; 102.4 ± 8.82 nmol/l). The inter-assay precision (CV%) was 11.9 (mean of 4 pools−7.03, 23.16, 52.11 and 117.53 nmol/l over 2 months). The mean bias was −0.78% over the range 28 to 448 nmol/l and different aliquots (equivalent to 0.25 to 4 μl) of daily early morning urine gave results that were in good agreement. The concentration (nmol/l; mean ± sd) of oestrone-3-glucuronide in samples of early morning urine collected from healthy women during the early follicular, periovulatory and luteal phases of the menstrual cycle were 40.2 ± 9.9, 102.3 ± 39.4 and 84.3 ± 13.3 nmol/l, respectively. In addition, the results obtained from the analysis of EMU throughout 6 complete menstrual cycles were in good agreement with the values derived by radioimmunoassay (r = 0.94).

Direct chemiluminescence immunoassay for estradiol in serum.

1986 ◽  
Vol 32 (10) ◽  
pp. 1895-1900 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
C Usanachitt ◽  
D Vandekerckhove ◽  
D Leyseele ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
High Affinity ◽  
Microtiter Plates ◽  
Analytical Recovery

Abstract In this simple, reliable, fast solid-phase chemiluminescence immunoassay for directly measuring (i.e., without prior extraction) estradiol-17 beta in serum, a monoclonal antibody is used that binds estradiol with high affinity (Ka = 10(10) L/mol), and does not bind other steroids tested, the highest cross reactivity observed being 0.1% for estradiol-17 alpha. In this system the monoclonal antibody is bound to the wells of microtiter plates via a second antibody directed against the monoclonal antibody. Fifty microliters of serum and estradiol-displacing agents are added, followed by 100 pg of estradiol-isoluminol conjugate, and the label is measured by luminometry after the binding reaction. The sensitivity of the assay is 180 pmol per liter of serum, and the effective working range at less than or equal to 10% CV is 270 to 6700 pmol/L. Analytical recovery of added estradiol averaged 99.7% (SD 6.5%). Within- and between-assay CVs ranged between 5 and 12.7%. Thirty-five unknown serum samples can be assayed within 4 h. Results correlated well with those obtained with a direct RIA: r = 0.94 (n = 149). This assay opens new perspectives for chemiluminescence immunoassays.

Development of a solid-phase chemiluminescence immunoassay for plasma progesterone

Steroids ◽  
1981 ◽  
Vol 38 (1) ◽  
pp. 73-88 ◽  
Author(s):  
F. Kohen ◽  
J.B. Kim ◽  
H.R. Lindner ◽  
W.P. Collins
Keyword(s):  
Solid Phase ◽  
Chemiluminescence Immunoassay ◽  
Plasma Progesterone

The Measurement of Urinary LH, by a Solid-Phase Chemiluminescence Immunoassay

1984 ◽  
Vol 21 (4) ◽  
pp. 284-289 ◽  
Author(s):  
J L Brockelbank ◽  
J B Kim ◽  
G J Barnard ◽  
W P Collins ◽  
B Gaier ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Sodium Hydroxide ◽  
Solid Phase ◽  
Early Morning ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Time Interval ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
Menstrual Cycles

Ovulation in women may be predicted or detected by a simple, solid-phase, chemiluminescence immunoassay for the measurement of urinary LH. An IgG fraction of a donkey antiserum to rabbit immunoglobulins is passively adsorbed to the walls of polystyrene tubes. Human chorionic gonadotrophin-succinyl-butyl-ethyl-isoluminol and a primary antibody (rabbit anti-human LH) is incubated with samples of early morning urine. After the binding reaction, the solution is removed by aspiration. The antibody-bound fraction is washed twice with buffer. Sodium hydroxide is added and the mixture incubated. After cooling, luminescence is initiated by oxidation of the label with microperoxidase/hydrogen peroxide and the signal integrated. The method has been evaluated in terms of sensitivity, within- and between-batch precision, bias and parallelism. The time interval between the peak day of LH in EMU and maximum follicular diameter during 38 menstrual cycles was −24 h (11%), 0 h (50%), + 24 h (28%) and + 48 h (11%).

Solid-phase chemiluminescence immunoassay for progesterone in unextracted serum.

1984 ◽  
Vol 30 (10) ◽  
pp. 1637-1641 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
D Vandekerckhove ◽  
G Van Maele
Keyword(s):  
Binding Proteins ◽  
Solid Phase ◽  
Free Ligand ◽  
Chemiluminescence Immunoassay ◽  
Binding Reaction ◽  
Direct Assay ◽  
Precision And Accuracy ◽  
Sensitivity Specificity

Abstract We describe a simple, solid-phase chemiluminescence immunoassay for progesterone in 10 microL of unextracted serum ("direct" assay). Danazol at pH 8.0 is included (100 ng per tube) to displace progesterone from binding proteins in serum. A progesterone-11 alpha-hemisuccinyl-aminobutylethyl isoluminol conjugate serves as the chemiluminescent ligand marker and homologous antiprogesterone IgG covalently coupled to "Immunobeads" is the immunoadsorbant. After the binding reaction, bound and free ligand are separated by centrifugation and the chemiluminescence yield of the bound label is determined. The sensitivity, specificity, precision, and accuracy of the method are similar to those of a conventional radioimmunoassay for progesterone in which a radioligand of tritiated progesterone and serum extraction are used. Progesterone values obtained by this procedure agreed well (r = 0.987) with those obtained by radioimmunoassay. We conclude that the chemiluminescence immunoassay for progesterone in unextracted serum is analytically valid and offers a convenient alternative to radioimmunoassay.

PROGESTERONE IN THE HUMAN PERIPHERAL BLOOD IN THE PREOVULATORY PERIOD OF THE MENSTRUAL CYCLE

1967 ◽  
Vol 55 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Benno Runnebaum ◽  
Josef Zander
Keyword(s):  
Menstrual Cycle ◽  
Corpus Luteum ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Progesterone Concentration ◽  
Plasma Progesterone ◽  
Progesterone Secretion ◽  
Graafian Follicle ◽  
The Mean ◽  
Mean Concentration

ABSTRACT Progesterone was determined and identified in human peripheral blood during the preovulatory period of the menstrual cycle, by combined isotope derivative and recrystallization analysis. The mean concentration of progesterone in 1.095 ml of plasma obtained 9 days before ovulation was 0.084 μg/100 ml. However, the mean concentration of progesterone in 1.122 ml of plasma obtained 4 days before ovulation was 0.279 μg/100 ml. These data demonstrate a source of progesterone secretion other than the corpus luteum. The higher plasma-progesterone concentration 4 days before ovulation may indicate progesterone secretion of the ripening Graafian follicle of the ovary.

scholarly journals Expression Profiles of the Progesterone Receptor, Cyclooxygenase-2, Growth Differentiation Factor 9, and Bone Morphogenetic Protein 15 Transcripts in the Canine Oviducts during the Oestrous Cycle

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 454
Author(s):  
Jaime Palomino ◽  
Javiera Flores ◽  
Georges Ramirez ◽  
Victor H. Parraguez ◽  
Monica De los Reyes
Keyword(s):  
Gene Expression ◽  
Bone Morphogenetic Protein ◽  
Cyclooxygenase 2 ◽  
Oestrous Cycle ◽  
Progesterone Concentration ◽  
Plasma Progesterone ◽  
Morphogenetic Protein ◽  
Growth Differentiation Factor 9 ◽  
Bone Morphogenetic Protein 15 ◽  
Cox 2

The gene expression in the canine oviduct, where oocyte maturation, fertilization, and early embryonic development occur, is still elusive. This study determined the oviductal expression of (PR), cyclooxygenase-2 (COX-2), growth differentiation factor 9 (GDF-9), and bone morphogenetic protein 15 (BMP-15) during the canine oestrous cycle. Samples were collected from bitches at anoestrus (9), proestrus (7), oestrus (8), and dioestrus (11), after routine ovariohysterectomy and the ovarian surface structures and plasma progesterone concentration evaluated the physiological status of each donor. The oviductal cells were isolated and pooled. Total RNA was isolated, and gene expression was assessed by qPCR followed by analysis using the t-test and ANOVA. The PR mRNA increased (P < 0.05) from the anoestrus to dioestrus with the plasma progesterone concentration (r = 0.8). COX-2 mRNA expression was low in the anoestrus and proestrus, and negligible in the oestrus, while it was around 10-fold higher (P < 0.05) in the dioestrus. The GDF-9 mRNA was expressed during all phases of the oestrous cycle and was most abundant (P < 0.05) during oestrus phase. The BMP-15 mRNA decreased (P < 0.05) in the anoestrus and proestrus phases. Thus, the transcripts were differentially expressed in a stage-dependent manner, suggesting the importance of oestrous cycle regulation for successful reproduction in dogs.

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