Akzo N.v.

Per Immune Holdings, Inc.

Ray F Ebert

Richard D Swerdlow

Rafael P Valdes-Camin

The present invention is directed to a stabilized keyhole limpet hemocyanin (KLH) composition in which (i) its intact non-degraded subunit is approximately 400,000 in molecular weight based on SDS-PAGE analysis; and (ii) are contained at least about 50% didecameric or higher KLH multimers, based on sedimentation-equilibrium and/or sedimentation-velocity ultracentrifugation analyses. The KLH composition is stabilized at 4.degree. C. by dissolving and storing it in an isotonic buffer preferably containing calcium and magnesium. It is critical that the KLH not have been frozen or lyophilized during its preparation or storage. The KLH composition demonstrates enhanced immunogenic activity, particularly enhanced anti-tumor activity, which is reduced if the KLH is frozen or lyophilized. The KLH composition of the present invention exhibits enhanced anti-tumor activity in a murine bladder tumor model and thereby represents a new and useful anti-tumor immunotherapeutic agent.

Method for treating bladder cancer with a keyhole limpet hemocyanin

The present intention is directed to a stabilized keyhole limpet hemocyanin (KLH) composition in which (i) its intact non-degraded subunit is approximately 400,000 in molecular weight based on SDS-PAGE analysis; and (ii) are contained at least about 50% didecameric or higher KLH multimers, based on sedimentation-equilibrium and/or sedimentation-velocity ultracentrifugation analyses. The KLH composition is stabilized at 4.degree. C. by dissolving and storing it in an isotonic buffer preferably containing calcium and magnesium. It is critical that the KLH not have been frozen or lyophilized during its preparation or storage. The KLH composition demonstrates enhanced immunogenic activity, particularly enhanced anti-tumor activity, which is reduced if the KLH is frozen or lyophilized. The KLH composition of the present invention exhibits enhanced anti-tumor activity in a murine bladder tumor model and thereby represents a new and useful anti-tumor immunotherapeutic agent.

Method using keyhole limpet hemocyanin composition with enhanced

A method for preparing a thromboplastin reagent from cultured human cells comprising the steps of washing the cells with isotonic aqueous salt solution, lysing the cells by hypotonic shock, and resuspending the cell lysate in a diluent compatible with a one-stage prothrombin time determination to reach a concentration of about 0.5 to 3.0 mg/mL thromboplastin, whereby the cell lysate thromboplastin suspension produces a clot in a one stage prothrombin time test in about 10 to 15 seconds, with a mean normal prothrombin time of about 11-13 seconds, when added to citrated normal plasma in a volume ratio of about 2:1, such thromboplastin reagent being stable for at least 8 hours at 37.degree. C. and for at least 5 days at 4.degree. C.

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Derwood, MD, United States of America

Ray F Ebert