Published October 1978 | Version v1
Report Open

Critical evaluation of the radioiodination of follicle-stimulating-and luteinizing hormones by lactoperoxidase and its comparison with chloramine-T classical method

Description

A method is described for the enzymatic radioiodination of human gonadotropins by a system consisting of lactoperoxidase, hydrogen peroxide and Na125I. A comparison witth the clhloramine-T modified technique was done a satisfactory specific activity (100 μCi/μg) of the labeled hormone was obtained with the enzymatic iodination, with much greater immunoreactivity and stability than after chloramine-T. As aqueous polyethylene glycol causes precipitation of antibody-bound peptide hormones with radioactive iodine with little or no precipitation of free hormones, a method of separation was developed and applied to radioimmunoassay of gonadotropins, providing several advantages over te double-antibody precipitation method. It was demonstrated that, in humans, the intravenous administration of synthetic LH-FSH/RH, stimulates the pituitary release of both LH and FSH. Results are reported now for normal women, infused with an acute load of 25μg of synthetic LH/FSH-RH and 8 hours infusion of 100μg, during the mid-follicular and luteal phases. The greatest gonadotropin responsiveness to LH/FSH-RH are found in the luteal phase during acute testing. No differences were noticed during chronic infusion as well as acute post-chronic, performed in both phases of the menstrual cycle. The possible modulating effects of steroidal secretion by the ovaries are discussed. (Author)

Availability note (English)

MF available from INIS under the Report Number.

Abstract (Portuguese)

A comparacao entre as duas tecnicas de marcacao das gonadotrofinas hipofisarias humanas, lactoperoxidase e cloramina-T, mostrou a vantagem da utilizacao do metodo enzimatico na reacao de substituicao de 125I ao radical tirosila. A oxidacao quimica com cloramina-T, apresentou o inconveniente da degradacao molecular, bem como a diminuicao da imunorreatividade desses hormonios glicoproteicos. Apos obtida a gonadotrofina 125I, desenvolveu-se a tecnica do polietilenoglicol (PEG) 4000 a 16%, em substituicao ao metodo classico de precipitacao com o segundo anticorpo, para separacao do hormonio livre daquele ligado ao anticorpo. Estudou-se a liberacao de LH e FSH hipofisarios pelo fator liberador das gonadotrofinas (FLG), em oito mulheres normais, nas duas fases do ciclo menstrual. Fizeram-se duas provas, uma aguda (injecao de 25μ de FLG) e outra prolongada (infusao de 100μg de FLG por 8 horas) seguida de uma aguda, tanto na metade da fase folicular como da correspondente na fase lutea. Observou-se resposta significativamente maior do LH na fase lutea, havendo diferenca significativa na resposta do FSH nas duas fases do ciclo menstrual, na prova aguda isolada. Nas provas prolongada e aguda pos-cronica a resposta nao foi significantemente diferente para os dois hormonios, nas duas fases do ciclo menstrual. Discutem-se os possiveis efeitos moduladores dos esteroides sexuais. (Autor)

Files

11552268.pdf

Files (910.6 kB)

Name Size Download all
md5:def77bf1be41bca063446f544a989865
910.6 kB Preview Download

Additional details

Additional titles

Subtitle (English)
Application in the seric gonadotropin assay in menstrual cycle after stimulation by hypothalamic liberator factor
Original title (Portuguese)
Avaliacao critica da radioiodacao dos hormonios luteinizante e foliculo estimulante hipofisario humanos com lactoperoxidase e sua comparacao com o metodo classico da cloramina-T
Original subtitle (Portuguese)
Aplicacao na medida das gonadotrofinas sericas, no ciclo menstrual, apos estimulo com fator liberador hipotalamico.

Publishing Information

Imprint Pagination
41 p.
Journal Issue
no. 085
Series
IEA Dissertacao e Tese.
Report number
IEA-DT--085