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Immunophototherapy of cancer combines the specificity of a monoclonal antibody (MAb) to an overexpressed tumor marker with the phototoxic properties of the conjugated dye. To analyze the potential role of internalisation of the dye on photo-induced cytotoxicity, we compared two target antigens, carcinoembryonic antigen (CEA) that does not internalise and ErbB2 that does. Human ovarian carcinoma SKOv3 cells that express a high level of ErbB2 were transfected with the CEA cDNA. Using FACS analysis, the resulting cell line, SKOv3-CEA-1B9, demonstrated comparable levels of expression of the two target antigens. Aluminium tetrasulfophthalocyanine (AlPcS 4) was covalently coupled to anti-CEA MAb 35A7, anti-ErbB2 MAb FSP77 and a non-specific MAb PX, via a five-carbon sulfonamide spacer chain (A 1) at molar ratios ranging from 6 to 9 moles of AlPcS 4 per mole of MAb. The 35A7-(AlPcS 4 A 1)8 conjugate induced 68% growth inhibition of the SKOv3-CEA-1B9 cell line after a 20 h incubation at 2.50 μg/ml (based on AlPcS 4 A 1 content) following light exposure. However, the FSP77-(AlPcS 4 A 1)6 conjugate gave a 51% growth inhibition for an AlPcS 4 A 1 concentration as low as 0.04 μg/ml after the same incubation time and exposure to the same light dose. At a 1.25 μg/ml AlPcS 4 A 1 concentration, the FSP77-(AlPcS 4 A 1)6 conjugate gave a 67% growth inhibition after an incubation time as short as 1 h, reaching a 96% inhibition after an 8 h incubation time. Using an unique cell line that expresses two different target antigens, we demonstrated a clear advantage of an internalising over a non-internalising MAb-dye conjugate in terms of phototoxic efficacy. In vivo evaluation of the photodynamic properties of the conjugates is in progress. © 2001 Cancer Research Campaign http://www.bjcancer.com © 2001 Cancer Research Campaign
immunophototherapy, phthalocyanine, monoclonal antibody, CEA, ErbB2
Internalisation enhances photo-induced cytotoxicity of monoclonal antibody-phthalocyanine conjugates
M Carcenac,1 M Dorvillius,1 V Garambois,1 F Glaussel,1 Ch Larroque,2 R Langlois,3 N E Hynes,4 J E van Lier,3 and A Pelegrin1
Anti-Mullerian hormone (AMH) [also called Mullerian inhibiting substance (MIS)] is a member of the transforming growth factor-beta family. AMH and its type II receptor (AMHR-II) are involved in the regression of the Mullerian ducts in the male embryo, and in gonadal functions in the adult. AMH is also known to be a marker of granulosa and Sertoli cell tumours. We selected a high-affinity monoclonal antibody, mAb 12G4, specific for human AMHR-II (hAMHR-II), by FACS analysis, Western blotting and immunohistochemical staining of a hAMHR-II-transfected CHO (Chinese hamster ovary) cell line, normal adult testicular tissue and granulosa cell tumours. Using peptide array screening, we identified the binding sequences of mAb 12G4 and AMH on the receptor. Identification of Asp53 and Ala55 as critical residues in the DRAQVEM minimal epitopic sequence of mAb 12G4 definitively accounted for the lack of cross-reactivity with the murine receptor, in which there is a glycine residue in place of an aspartic acid residue. In a structural model, the AMH-binding interface was mapped to the concave side of hAMHR-II, whereas the mAb 12G4-binding site was located on the convex side. mAb 12G4, the first mAb to be raised against hAMHR-II, therefore has unique properties that could make it a valuable tool for the immunotargeting of tumours expressing this receptor.
The anti-Mullerian hormone type II receptor: insights into the binding domains recognized by a monoclonal antibody and the natural ligand.
Imed Salhi, Sylvie Cambon-Roques, Isabelle Lamarre, Daniel Laune, Franck Molina, Martine Pugniere, Didier Pourquier, Marian Gutowski, Jean-Yves Picard, Francoise Xavier, Andre Pelegrin, and Isabelle Navarro-Teulon
2004 May 1;
Metastatic breast cancer (MBC) prognosis is variable, depending on several clinical and biological factors. A better prediction of a patient’s outcome could allow for a more accurate choice of treatments. The role of serum biomarkers in predicting outcome remains unclear in this setting. Tau, a microtubule-associated protein, is a neuronal marker that is also expressed in normal breast epithelial cells and cancer cells. Its tissue expression is associated with prognosis in MBC. However, the prognostic value of Tau serum levels in these patients is unknown. We aimed at evaluating the prognostic value of Tau (and other classical biomarkers) in MBC patients, and to assess its association with the presence of brain metastases (BM).
244 MBC patients treated at our institution (2007-2015) were retrospectively selected. The usual MBC clinical and pathological variables were collected, altogether with CA15-3, CEA and HER2 extra-cellular domain (ECD) serum levels. Tau serum levels were measured with a novel immunoassay (digital ELISA) using Single Molecule Array (Simoa) technology. Overall survival (OS) was estimated with the Kaplan-Meier method. To investigate prognostic factors, a multivariate analysis was performed. Cut-offs were set using the Youden index method associated with receiver-operating characteristics (ROC) curves to evaluate the accuracy of biomarkers to identify patients with BM.
With a median follow-up of 40.8 months, median OS was 15.5 months (95%CI 12.4-20.2). Elevated serum levels of Tau were independently associated with a poor outcome in the whole population as well as in patients with (n = 86) and without BM (n = 158). Median serum Tau levels tended to be higher in patients with BM (p = 0.23). In univariate analysis, patients with BM had an increased risk of serum Tau > 3.17 pg/mL (OR = 2.2, p = 0.049). In multivariate analysis, high values of Tau (OR = 3.98, p = 0.034) accurately identified patients with BM in our cohort.
Tau is a new biomarker of interest in MBC. Its serum level could represent an independent prognostic factor in these patients (both with and without BM). It also seems to be associated with the presence of BM. A validation of these results in an independent set of MBC patients is necessary to confirm these findings.
Electronic supplementary material
The online version of this article (10.1186/s12885-019-5287-z) contains supplementary material, which is available to authorized users.
Tau protein, Breast cancer, Brain metastases, Tumor markers, Predictive factors, Prognostic factors
The prognostic value of the Tau protein serum level in metastatic breast cancer patients and its correlation with brain metastases
Amelie Darlix,corresponding author1 Christophe Hirtz,2 Simon Thezenas,3 Aleksandra Maceski,2 Audrey Gabelle,4 Evelyne Lopez-Crapez,5 Helene De Forges,6 Nelly Firmin,1 Severine Guiu,1 William Jacot,1 and Sylvain Lehmann2