Urinary gonadotrophin peptide – isolation and purification, and its immunohistochemical distribution in normal and neoplastic tissues

Kardana, Andrew; Taylor, Marisa; Southall, P. J.; Boxer, G. M.; Rowan, AJ; Bagshawe, K. D.

doi:10.1038/bjc.1988.204

Cited by 39 publications

(23 citation statements)

References 13 publications

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“…It has also been detected in 93% (77/83) of tumours examined immunohistochemically (Kardana et al, 1988). The apparent molecular weight is 15,000.…”

mentioning

confidence: 86%

“…A commercial preparation of hCG (Pregnyl) derived from pooled pregnancy urines was subjected to gel filtration on Sephadex G 100 and immunoadsorption with a mouse monoclonal P-directed antibody (W14) immobilised on cyanogen-bromide activated Sepharose 4B, the required material being eluted with 3 M ammonium thiocyanate followed by immediate desalting by G25 chromatography, according to procedures published earlier (Kardana et al, 1988). Aliquots of the immunoadsorbed material were further purified on FPLC using a Superose 12 column (Pharmacia FPLC systems) or re-subjected to gel-filtration using Sephadex G-100.…”

Section: Purification Of Ugpmentioning

confidence: 99%

“…UGP from normal male urine or ovarian carcinoma urine was extracted using acetone precipitation (as described in Kardana et al, 1988), whilst urine from a patient with hydatidiform mole was extracted using either acetone precipitation (as above) or as an alternative kaolin adsorption was used. Two litres of urine were adjusted to pH=4.5 with glacial acetic acid and then 35 g of kaolin (a modification of the method of Albert, 1956, that does not include the final precipitation step) stirred into this solution.…”

Section: Purification Of Ugpmentioning

confidence: 99%

“…Beta core-fragment on iso-electric focusing was found to have a pl > 9.5, peptide 3 showed two bands at pl = 3.5 and 3.8 whilst insufficient purified peptide 1 was available to determine its iso-electric point. Bioassay studies on UGP showed that any biological activity could be attributed to trace contamination with hCG.Urinary gonadotrophin peptide (UGP) has been purified from the urine of patients with trophoblastic and nontrophoblastic disease (Kardana et al, 1988). It has also been detected in 93% (77/83) of tumours examined immunohistochemically (Kardana et al, 1988).…”

mentioning

confidence: 99%

“…Urinary gonadotrophin peptide (UGP) has been purified from the urine of patients with trophoblastic and nontrophoblastic disease (Kardana et al, 1988). It has also been detected in 93% (77/83) of tumours examined immunohistochemically (Kardana et al, 1988).…”

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confidence: 99%

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Characterisation of UGP and its relationship with beta-core fragment

Kardana

Bagshawe²,

Coles³

et al. 1993

Br J Cancer

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Summary Urinary gonadotrophin peptide (UGP) was originally identified by immunoassay in the urine of patients with various types of cancer and by immunohistochemistry in human cancers of various histological types. Extracts of normal adult male urine also contained UGP by immunoassay. Purified UGP from different starting material was subjected to high pressure liquid chromatography (HPLC) prior to defining amino acid sequences. Chromatographed UGP after HPLC showed three distinct fractions. The N-terminal sequence of peptide 2 was completely homologous with the beta-core fragment of human chorionic gonadotrophin (hCG) and this was found associated with two smaller peptides. The N-terminal sequence of peptide has not been described previously whilst the N-terminus of peptide 3 that was sequenced showed complete homology with the N-terminal sequence of eosinophil derived neurotoxin and non-secretory ribonuclease. The monoclonal antibodies 2C2 and 6D3 only bind beta core-fragment (peptide 2) whilst the polyclonal (rabbit) antibody AK12 could bind all three peptides. The radioimmunoassay system using AK12 could be inhibited by all three peptides and the immunoradiometric assay although based on a capture antibody (2C2) that only bound peptide 2, had the potential to measure all three peptides (when bound together as UGP) at the second step when '25l-AK12 was introduced as the detector. A specific radioimmunoassay for peptide 3 was generated using 1251-peptide 3 and the AK12 antibody. Beta core-fragment on iso-electric focusing was found to have a pl > 9.5, peptide 3 showed two bands at pl = 3.5 and 3.8 whilst insufficient purified peptide 1 was available to determine its iso-electric point. Bioassay studies on UGP showed that any biological activity could be attributed to trace contamination with hCG.Urinary gonadotrophin peptide (UGP) has been purified from the urine of patients with trophoblastic and nontrophoblastic disease (Kardana et al., 1988). It has also been detected in 93% (77/83) of tumours examined immunohistochemically (Kardana et al., 1988). The apparent molecular weight is 15,000. UGP cross reacts with antisera to hCG beta-core fragment, which contains at least one epitope in common with the beta-subunit of hCG, as seen by the ability to measure beta-core fragment using antisera raised to hCG beta-subunit Papapetrou & Nicopoulou, 1986;Wehmann & Nisula, 1980). Initial screening of urines using specific immunoassays for UGP (Kardana et al., 1989) has shown detectable levels of UGP in normal subjects and elevated levels in the urine of some patients with neoplasms (manuscript submitted for publication).The origin of beta-core fragment (BCF) is unclear and there are reports that it is a renal breakdown product of either hCG or its beta-subunit . It has been shown that the major form of betacore fragment in serum exists as a high molecular weight complex (Mr >60,000) which can be dissociated with 3M ammonium thiocyanate to release beta-core fragment (Mr= 15,000) (Kardana & .In this paper we report further ...

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“…It has also been detected in 93% (77/83) of tumours examined immunohistochemically (Kardana et al, 1988). The apparent molecular weight is 15,000.…”

mentioning

confidence: 86%

Section: Purification Of Ugpmentioning

confidence: 99%

Section: Purification Of Ugpmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Characterisation of UGP and its relationship with beta-core fragment

Kardana

Bagshawe²,

Coles³

et al. 1993

Br J Cancer

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Urinary chorionic gonadotropin subunits and β-core in nonpregnant women. A study of benign and malignant gynecologic disorders

Neven

Iles

Lee

et al. 1993

Cancer

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Background. The presence of urinary excretion products of human chorionic gonadotropin (hCG) has been proposed as a tumor marker. To ascertain the clinical value in gynecologic cancers, the authors studied 612 nonpregnant women. Methods. Three different assays in four clinical groups were compared: no disease, benign disease, malignant disease, and complete remission of previously treated malignant disease. The assays were for the urinary β‐core, “total” β‐hCG, and free α‐subunit. Results. Measurement of the α‐subunit was of no obvious clinical value. In some patients with benign disease, hCG metabolites were elevated. In the 141 patients with active gynecologic malignancy the sensitivity of the total β‐hCG assay was 47% and that of the β‐core assay was 36%. The specificities were 80.3% and 90.4%, respectively. Advanced cancers generally had higher levels of total β‐hCG and β‐core. Squamous cell and poorly differentiated cervical tumors had higher levels of total β‐hCG than did adenocarcinomas and well‐differentiated cervical tumors. Invasive, serous, endometrioid, and germ cell ovarian tumors had higher total β‐hCG, β‐core, and α‐subunit levels than did borderline, mucinous, and clear cell ovarian tumors. Six of 16 patients with disease in complete remission had elevated levels. Conclusion. The excretion of hCG and its metabolic fragments is a common event in gynecologic cancer, but sensitivity and specificity are low, and there is little consistent relationship between tumor stage and histologic type.

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Urinary gonadotropin peptide: Collection of specimens and cutoff levels

Plebani

Navaglia

Bacelle

et al. 1995

Clinical Laboratory Analysis

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A low-molecular-weight form of human chorionic gonadotropin (hCG), urinary gonadotropin peptide (UGP), has been isolated from the urine of pregnant women and of patients with cancer, mainly of gynecological origin. The clinical value of UGP measurement in gynecological diseases is under investigation but a preliminary study is necessary in order to ascertain whether there is a circadian rhythm in UGP production, to clarify the best way to express the results, and to establish the cutoff and decisional values. In our work we demonstrated a significant correlation between the UGP output and the UGP excretion normalized for urinary creatinine. A very significant agreement was even found in 24-hr urine collections and UGP concentration of a single morning specimen from the same patients. No evident circadian rhythm was found, although some patients presented morning levels of UGP higher than in other collections. UGP postmenopausal levels were higher than premenopausal. The cutoff level, adopting the 95.0 percentile, was 200 pmol/mol creatinine.

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Urinary gonadotrophin peptide – isolation and purification, and its immunohistochemical distribution in normal and neoplastic tissues

Cited by 39 publications

References 13 publications

Characterisation of UGP and its relationship with beta-core fragment

Characterisation of UGP and its relationship with beta-core fragment

Urinary chorionic gonadotropin subunits and β-core in nonpregnant women. A study of benign and malignant gynecologic disorders

Urinary gonadotropin peptide: Collection of specimens and cutoff levels

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