Preparation and properties of a mouse monoclonal antibody (W14A) to human chorionic gonadotropin

Searle, F.; Partridge, Christine S.; Kardana, Andrew; Green, A. J.; Buckley, R. G.; Begent, R. H. J.; Rawlins, G.A.

doi:10.1002/ijc.2910330402

Cited by 15 publications

(13 citation statements)

References 17 publications

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“…Immunopurified mouse monoclonal W14A anti-human chorionic gonadotrophin and F (ab')2 fragments of the same antibody were produced by previously described methods (Searle et al, 1984(Searle et al, , 1986b by the Department of Medical Oncology, Charing Cross Hospital, London, UK.…”

Section: Methodsmentioning

confidence: 99%

“…'l3I-labelled W14A or F (ab')2 were produced by a modification of the same method in which the reduction of free iodide to iodine by sodium metabisulphite was replaced by the addition of L-tyrosine (100 jld of a freshly prepared solution, 2 mg ml-' in PBS). This modification to the method has been reported to result in less immunological deterioration of the antibody (Searle et al, 1984) Conjugates of radiolabelled CPG2 and W14A or its F (ab')2 fragment were prepared using MBS heterobifunctional reagent by previously described techniques (Searle et al, 1986a). The conjugates were purified by gel filtration chromatography on a column (2.2 x 90 cm) of Ultrogel AcA34, with PBS as elution buffer.…”

Section: Methodsmentioning

confidence: 99%

“…Bertino, personal communication). In view of this potential application and its use of an activator of pro-drugs, we have studied the persistence and fate of conjugates of CPG2 with anti-human chorionic gonadotrophin (anti-hCG) or its F (ab')2 fragment as a means of selectively delivering the enzyme to CC3 choriocarcinoma xenografts borne by nude mice and rats (Searle et al, 1981).…”

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confidence: 99%

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The potential of carboxypeptidase G2: antibody conjugates as anti-tumour agents. II. In vivo localising and clearance properties in a choriocarcinoma model

Melton

Searle²,

Sherwood³

et al. 1990

Br J Cancer

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Summary The in vivo localising and clearance properties of conjugates of the folate-degrading enzyme carboxypeptidase G2 (CPG2) with anti-human chorionic gonadotrophin (W14A) were measured in nude mice bearing CC3 choriocarcinoma xenografts. Conjugates of W14A-F (ab')2 fragment coupled to CPG2 localised in tumour as effectively as native antibody alone but showed lower uptake in other major tissues. The clearance rates of conjugates prepared with intact antibody or F (ab')2 fragment were shown to be up to five-fold faster than for native antibody and two-fold compared to F (ab')2 fragment. Molecular weight analysis of residual conjugate in the blood showed that no degradation of conjugate to its component molecules occurred during circulation. It was concluded that F (ab')2: CPG2 conjugates offered the greatest potential for targeting applications.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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The potential of carboxypeptidase G2: antibody conjugates as anti-tumour agents. II. In vivo localising and clearance properties in a choriocarcinoma model

Melton

Searle²,

Sherwood³

et al. 1990

Br J Cancer

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“…The only antibody preparation which showed any binding with peptides 1 and 3 as well as peptide 2 was the AK12 polyclonal (rabbit) antiserum. Further details on epitope specificities have been published earlier (Kardana et al, 1988, Kardana, 1990Searle et al, 1984).…”

Section: Antibody Binding Datamentioning

confidence: 99%

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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“…The mouse monoclonal antibody (W14), an IgG1 antihuman chorionic gonadotrophin (hCG), was produced either in mouse ascites or cell free supernatant and immunopurified on an hCG-sepharose column as previously described (Searle et al, 1984). Ultracentrifugation was performed after the method of Rossen et al (1971).…”

Section: Monoclonal Antibodymentioning

confidence: 99%

Cyclosporin A prevents the anti-murine antibody response to a monoclonal anti-tumour antibody in rabbits

Ledermann

Begent²,

Bagshawe³

1988

Br J Cancer

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Preparation and properties of a mouse monoclonal antibody (W14A) to human chorionic gonadotropin

Cited by 15 publications

References 17 publications

The potential of carboxypeptidase G2: antibody conjugates as anti-tumour agents. II. In vivo localising and clearance properties in a choriocarcinoma model

The potential of carboxypeptidase G2: antibody conjugates as anti-tumour agents. II. In vivo localising and clearance properties in a choriocarcinoma model

Characterisation of UGP and its relationship with beta-core fragment

Cyclosporin A prevents the anti-murine antibody response to a monoclonal anti-tumour antibody in rabbits

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