New assays for preimplantation factor and preimplantation...

C - Chemistry – Metallurgy – 07 – K

Patent

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Details

C07K 14/47 (2006.01) C07K 7/06 (2006.01) C07K 7/08 (2006.01) G01N 33/566 (2006.01) G01N 33/567 (2006.01)

Patent

CA 2490538

The present invention relates to assay methods used for detecting the presence of PIF, and to PIF peptides identified using this assay. In particular, the present invention relates to flow cytomery assays for detecting PIF. It is based, at least in part, on the observation that flow cytometry using fluorescently labeled antilymphocyte and anti-platelet antibodies demonstrated an increase in rosette formation in the presence of PIF. It is further based on the observation that flow cytometry demonstrated that monoclonal antibody binding to CD2 decreased in the presence of PIF. The present invention further relates to PIF peptides which, when added to Jurkat cell cultures, have been observed to either (I) decrease binding of anti-CD2 antibody to Jurkat cells; (ii) increase expression of CD2 in Jurkat cells; or (iii) decrease Jurkat cell viability. In additional embodiments, the present invention provides for ELISA assays which detect PIF by determining the effect of a test sample on the binding of anti-CD2 antibody to a CD2 substrate.

La présente invention concerne des méthodes de dosage utilisées pour détecter la présence de PIF; et des peptides de PIF identifiés au moyen de ce dosage. Elle concerne en particulier des dosages par cytométrie de flux destinés à détecter PIF. La méthode est fondée, au moins en partie, sur l'observation selon laquelle la cytométrie de flux utilisant des anticorps antilymphocytaires et antiplaquettaires marqués par fluorescence a révélé que l'anticorps monoclonal lié au CD2 diminuait en présence de PIF. L'invention concerne également des peptides de PIF qui, lorsqu'ils sont ajoutés à des cultures de cellules Jurkat, sont réputés soit i) diminuer la liaison de l'anticorps anti-CD2 aux cellules Jurkat; soit ii) augmenter l'expression de CD2 dans les cellules Jurkat; soit encore iii) diminuer la viabilité des cellules Jurkat. Dans une autre forme de réalisation, l'invention concerne des dosages ELISA pouvant détecter PIF par détermination de l'effet d'un échantillon d'essai sur la liaison de l'anticorps anti-CD2 à un substrat CD2.

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