Antiparathyroid antibodies which specifically bind to cell surface molecules on parathyroid, renal and cytotrophoblast cells which are involved in calcium sensing
OwnerClaes Juhlin, Rikard Holmdahl +3inventors
InventorsClaes Juhlin, Rikard Holmdahl, Lars Klareskog, Jonas Rastad, Goran Akerstrom
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Abstract
Antibody preparations are useful in the prognosis of parathyroid and renal disorders. The antibody preparation includes antibodies which bind specifically to a surface antigen, Ca.sup.2+ sensing structure, present in humans on the parathyroid, on the proximal tubuli of the kidney, on the cytotrophoblastic cells of the placenta, on renal carcinoma cells and on parathyroid cancer cells.
Claims6
A monoclonal antibody selected from the group consisting of B6 and G11, wherein the monoclonal antibody binds specifically to cell surface molecules present on renal, parathyroid and cytotrophoblastic cells, and wherein said surface molecules are involved in a Ca.sup.2+ sensing mechanism.
An antibody preparation selected from the group consisting of antibodies B6, G11 and mixtures thereof, which binds specifically to cell surface molecules present on renal, parathyroid and cytotrophoblast cells, wherein said antibody preparation binds to said surface molecules and affects a complete concentration-dependent abolishment of cytoplasmic Ca.sup.2+ increase when extracellular Ca.sup.2+ is increased, produced by a process comprising the steps of:
- a) producing hybridomas of antibodyproducing cells fused with myeloma cells;
- b) conducting immunohistochemical screening of hybridomas of step a) and selecting hybridomas with positive antibody activity specific for normal parathyroid cells;
- c) subcloning hybridomas with positive antibody activity of step b) which are specific for surface molecules of parathyroid cells;
- d) conducting immunofluorescence staining of subcloned hybridomas with positive antibody activity obtained in step c) to confirm that antibodies produced by said subcloned hybridomas give rise to distinct staining on surface structures of parathyroid cells;
- e) determining the isotypes of antibodies produced by said subcloned hybridomas by ELISA;
- f) purifying the antibodies;
- g) determining the effects of the antibodies on cytoplasmic calcium of parathyroid cells by microfluorimetrically recording individual cells with loading of fura-2;
- h) monitoring internal and external calcium with and without the antibodies to screen for those antibodies which block epitopes of the cells involved with the uptake of calcium, and
- i) screening said antibodies for.ability to bind renal and cytotrophoblast cells.
An antibody that binds to the cell surface molecules recognized by the B6 and G11 antibodies, wherein said cell surface molecules are present on renal, parathyroid and cytotrophoblast cells, and are involved in a Ca.sup.2+ sensing mechanism.
Dependencies are inferred from the claim text.