Use of monoclonal antibodies to pregnanediol-3α-glucuronide for the development of a solid phase chemiluminescence immunoassay

Eshhar, Zelig; Kim, J.B.; Barnard, Geoff; Collins, William P.; Gilad, S.; Lindner, H. R.; Konen, Felix Franz

doi:10.1016/0039-128x(81)90023-4

Cited by 53 publications

(9 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein A affinity chromatography, SDS-polyacrylamide gel electrophoresis, and reaction with rabbit anti-mouse p-chain antibody established that clone 107 produced an IgG. Monoclonal antibodies generated against covalent steroidal protein conjugates have generally been focnd to be of the IgG class (22,23,24,25) with a few of the IgM class (24,25). The general success rate in obtaining hybridoma cell lines synthesizing antibodies directed against steroids employing covalent conjugates are roughly equivalent to our results employing a noncovalent conjugate (22,23,24,25).…”

Section: Discussionsupporting

confidence: 80%

See 1 more Smart Citation

Production of monoclonal antibodies to a steroid plant growth regulator

Horgen¹,

Nakagawa²,

Irvin³

1984

Can. J. Biochem. Cell Biol.

View full text Add to dashboard Cite

1984) Production of monoclonal antibodies to a steroid plant growth regulator.Can. J . Biochem. Cell Biol. 62, [715][716][717][718][719][720][721] The active component of the brassin complex of higher plants has been shown to be a C28 steroidal lactone called brassinolide. This compound has been shown to enhance both cell division and cell growth. Monoclonal antibodies directed against a synthetic brassinosteroid have been produced in CAFB mice. The mice were immunized with a saline solution containing the steroid hapten noncovalently bound to chloroform-methanol extracted fetal calf serum. Hybridomas were generated by fusing primed mouse spleen cells with SP2/0 myeloma cells. Sixty-one hybridoma clones were screened; four of these gave positive enzyme-linked immunosorbent assay (ELISA) values with the brassinosteroid hapten. One of these clones produced an immunoglobulin G antibody with a high specificity for the synthetic antigen. The brassinosteroid hapten which differs from the natural brassinolide by the position of a methyl group on the C24 carbon gave a considerably higher ELISA value than the natural growth regulator and several other plant steroids. The distribution of brassinolide in Brassica napus tissues was examined by ELISA with the monoclonal antibody.Horgen, P. A., Nakagawa, C. W. & b i n , R. T. (1984) Production of monoclonal antibodies to a steroid plant growth regulator.Can. J . Biochem. Cell Biol. 62, 715-721 Le constituant actif du complexe brassin des plantes sup6rieures est une lactone stCroi'de comportant 28 atomes de carbsne, appelCe brassinolide. Ce compose favorise la division cellulaire et la croissance cellulaire. Nous avons produit des anticorps monoclonaux dirigCs contre un brassinsstCroi'de synthCtique dans les souris CAF,. Les souris Ctaient immunisCes avec une solution saline contenant 19hapt&ne sttrside lit de fason non covalente au serum de veau foetal extrait avec un melange de chlorofome-mCthano1. Des hybridomes sont gtndrts par fusion de cellules de rates de souris immunisdes avec les cellules du mytlome SP2/0. Nous avons examine 61 clones d9hybridomes; quatre de ces clones donne des valeurs ELZSA (test immunoenzymatique) positives avec l'hapdne brassinostQoide. L'un de ces clones produit un anticorps immunoglobuline C avec une spCcificitC ClevCe pour l'antigkne synthetique. L'hapthe brassinosdroi'de, qui diff&re du brassinolide naturel par la presence d'un groupe mkthyle sur le carbsne 24, donne une valeur ELISA considCrablement plus Clevie que le rCgulateur naturel de la croissance et plusieurs autres stCroi'des vtgetaux. Nous avons examine la distribution du brassinolide dans les tissus de Brassica napus par l'essai ELISA avec l'anticorps monoclonal.[Traduit par ]la revue]

show abstract

Section: Discussionsupporting

confidence: 80%

“…Although our data does not demonstrate complete specificity, it is generally cornparable to the specificity reported in other monclonal steroid studies (22,23,24,27,28).…”

Section: Discussionsupporting

confidence: 59%

Production of monoclonal antibodies to a steroid plant growth regulator

Horgen¹,

Nakagawa²,

Irvin³

1984

Can. J. Biochem. Cell Biol.

View full text Add to dashboard Cite

show abstract

“…Since that time several homogeneous and heterogeneous immunoassays have been reported for the measurement of ste¬ roids and their urinary metabolites using isoluminol conjugates (Kohen et al 1979(Kohen et al , 1980a(Kohen et al ,b, 1981aBarnard et al 1981a,b;Eshhar et al 1981;Kim et al 1982). The labelled antigens have been shown to be very stable over 2 years and the detection systems are relatively inexpensive and reliable.…”

Section: Discussionmentioning

confidence: 97%

“…Recently, chemiluminescent immunoassays have been developed for the measurement of urinary steroid metabolites (Barnard et al 1981a,b;Eshhar et al 1981) which have involved the use of a solid-phase separation system. The specific IgG is adsorbed onto the surface of polystyrene assay tubes and after the binding reaction the incubation mixture is aspirated.…”

mentioning

confidence: 99%

Monitoring ovarian function by a solid-phase chemiluminescence immunoassay

Weerasekera¹,

Kim²,

Barnard³

et al. 1982

Acta Endocrinologica

View full text Add to dashboard Cite

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\x=req-\ 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 \g=m\lremoved, in duplicate, for assay. After the binding reaction (1 h at 4\s=deg\C), the solution is removed by aspiration. The antibody-bound fraction is washed once with buffer (400 \ g=m\ l ). Sodium hydroxide (2n, 200 \g=m\l ) is added and the mixture incubated for 60 min at 22\s=deg\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 \m=+-\0.75 pg/tube (mean \m=+-\ sd). The intra-assay precision (CV%) was 9.52 (20 replicates; 38.4 \ m=+-\3.66 nmol/l) and 8.61 (15 replicates; 102.4 \ m=+-\ 8.82 nmol/l). The inter-assay precision (CV%) was 11.9 (mean of 4 pools\m=-\7. 03,23.16, 52.11 and 117.53 nmol/l over 2 months). The mean bias was \m=-\0.78% over the range 28 to 448 nmol/l and different aliquots (equivalent to 0.25 to 4 \g=m\l ) of daily early morning urine gave results that were in good agreement. The concentration (nmol/l; mean \ m=+-\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 \m=+-\9.9, 102.3 \ m=+-\ 39.4 and 84.3 \m=+-\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).Numerous methods have been devised to investi¬ gate ovarian function and to monitor or predict variations in potential fertility in the human female. These have been based on: i) the secretion of pituitary and ovarian hormones (WHO 1980); ii) their biochemical or biophysical effect on re¬ sponsive tissue (Moghissi 1980); or iii) the change in ovarian morphology as observed with ultrasound (Queenan et al. 1980). Consequently, a greater understanding of the interaction and temporal relationships of the many parameters associated with ovulation has been achieved.The results from studies of ovarian function and steroid metabolism have suggested that the prin¬ cipal urinary metabolites of oestradiol and proge¬ sterone might be used as indices of the start and finish of the probable fertile period ). Accordingly, methods based upon the prin¬ ciples of radioimmunoassay have been developed to measure the main oestrogen glucuronides and pregnanediol-3a-glucuronide in diluted urine (Samarajeewa & Kellie 1975; Samarajeewa et al. 1979). Subsequently, several investigators have undertaken the study of facets of this bochemical approach...

show abstract

“…1). C57/BL6 mice 18,19 and SD rats 20 were also immunized with the same immunogen, considering previous reports using these strains for successful production of monoclonal anti-steroid antibodies. However, no significant difference in the response was observed between these strains, and we selected two BALB/c mice and two A/J mice that showed a relatively high immune response as spleen donors.…”

Section: Production Of Monoclonal Antibodiesmentioning

confidence: 99%