Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1995-02-10
1996-11-19
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
53038827, 53038825, G01N 3353, C07K 1618
Patent
active
055761836
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a monoclonal antibody recognizing a protein which binds with FK506 (FK506 -binding protein) having an immunosuppressive activity, a method for assaying the level of an FK506-binding protein and a kit therefor, which are usable in the medical field.
BACKGROUND ART
A compound designated as FK506 or FR-900506 is well known to have a potent immunosuppressive activity and is usable as an agent for the prophylaxis and treatment of rejection by organ transplantation and autoimmune diseases (for example, Japanese Patent Unexamined Publication No. 14818/1986).
However, the activity is so potent that the determination of the optimal dose is of utmost importance and administration of an amount capable of inducing effective immunosuppressive activity without side effects is extremely important.
Later studies revealed that the immunosuppressive action of FK506 is caused by the binding with an intracellular FK506 -binding protein (hereinafter indicated as FKBP) having similar activity to peptidylprolyl cis-trans isomerase [for example, Nature, 357, 692-694 and 695-697 (1992)].
FKBP is known to include several types depending on the molecular weight, such as FKBP-12 (molecular weight 12 KDa), FKBP-13 (molecular weight 13 KDa), FKBP-25 (molecular weight 25 KDa), FKBP-56 (molecular weight 56 KDa), FKBP-80 (molecular weight 80 KDa) and the like, and their structures have been already identified.
See, for example, Nature, 341, 755-757 and 758-760 (1989), J. Am. Chem. Soc. 113, 1409-1411 (1991), Proc. Natl. Acad. Sci., 88, 6229-6233 (1991) and the like.
Also, the production of an FKBP having a molecular weight of 11,819 daltons and 107 amino acids by genetic engineering has been already reported [Nature, 346, 671-674 (1990)].
It has also been reported that FK506 most strongly binds with FKBP-12 from among those FKBPs and its affinity constant (Kd) is 0.4 nM, and that FKBP-12 is present in large amounts not only in lymphocytes but also in various tissues inclusive of erythrocytes [Trans. Proc. 23 (6), 2760-2762 (1991)].
In view of the fact that, when FKBP-12 is given to mouse MLR system together with a certain amount of FK506, the immunosuppressive effect of FK506 is inhibited in proportion to the amount of the FKBP-12 added, FK-506 will be bound with FKBP-12 in plasma if it is present in blood, and the immunosuppressive effect cannot be obtained to the degree expected from the concentration of FK506 in plasma. Considering that erythrocytes of patients who underwent operation lyse and FKBP-12 in erythrocytes circulates through plasma, and that patients show different sensitivities, there has been a demand for the development of a technique which enables precise assay of FKBP level in plasma, whereby patient's sensitivity to FK506 is presumed and more desirable FK506 level in plasma can be determined.
DISCLOSURE OF THE INVENTION
It is therefore an object of the present invention to provide a monoclonal antibody recognizing FKBP, a method for precisely assaying the level of FKBP in plasma for the determination of desirable FK506 level in plasma, and a kit to be used for the assay.
The present inventors have conducted intensive studies and succeeded in producing a monoclonal antibody which recognizes an antigenic determinant in FKBP and does not inhibit the binding between FKBP and FK506, and established a method for precisely assaying the level of FKBP in plasma and a kit for the assay.
Accordingly, the present invention relates to a monoclonal antibody which recognizes an antigenic determinant in FKBP and does not inhibit the binding between FKBP and FK506, a method for assaying an FKBP level, comprising reacting an immobilized monoclonal antibody with an enzyme-labelled FK506 and FKBP in a specimen, a method for assaying an FKBP level, comprising reacting an immobilized FKBP with an enzyme-labelled FK506 and FKBP in a specimen, and a kit for FKBP level assay, comprising said monoclonal antibody, FKBP and an enzyme-labelled FK506.
The monoclonal antibody which recognizes a
REFERENCES:
patent: 4894366 (1990-01-01), Okuhara et al.
Nature, vol. 357, Jun. 25, 1992, Stephen J. O'Keefe, et al., "FK-506- And CsA-Sensitive Activation Of The Interleukin-2 Promoter By Calcineurin", pp. 692-694.
Nature, vol. 357, Jun. 25, 1992, Neil A. Clipstone, et al., "Identification Of Calcineurin As A Key Signalling Enzyme In T-Lymphocyte Activation", pp. 695-697.
Nature, vol. 341, Oct. 26, 1989, John J. Siekierka, et al., "A Cytosolic Binding Protein For The Immunosuppressant FK506 Has Peptidyl-Prolyl Isomerase Activity But Is Distinct From Cyclophilin", pp. 755-757.
Nature, vol. 341, Oct. 26, 1989, Matthew W. Harding, et al., "A Receptor For The Immunosuppressant FK506 Is A Cis-Trans Peptidyl-Prolyl Isomerase", pp. 758-760.
J. Am. Chem. Soc., 1991, vol.113, pp. 1409-1411, Heinz Fretz, et al., "Rapamycin And FK506 Binding Proteins (Immunophilins)".
Proc. Natl. Acad. Sci, USA, vol. 88, Jul. 1991, pp. 6229-6233, Thomas Hultsch, et al., "Immunophilin Ligands Demonstrate Common Features Of Signal Transduction Leading To Exocytosis Or Transcription".
Transplantation Proceedings, vol. 23, No. 6, Dec. 1991, pp. 2760-2762, J. E. Kay, et al., "Uptake Of FK 506 By Lymphocytes And Erythrocytes".
Nature, vol. 256, Aug. 7, 1975, G. Kohler, et al., "Continuous Cultures Of Fused Cells Secreting Antibody Of Predefined Specificity", pp. 495-497.
Seuier et al. Clin Chem 27:1797-1806, 1981.
Jayaraman et al. Journal Biol. Chem. 267:9474-7 1992.
Rosborough et al. Transpl. Proc. vol. 23:2890-93, 1991.
Tsudo et al. PNAS USA 83:9694-8 1986.
Tamuro et al. Transpl. Proc. 19:23-9, 1987.
Callebaut et al, Proc. Natl. Acad. Sci. USA, vol. 89, pp. 6270-6274 (1992).
Lebeau et al, The Journal of Biological Chemistry, vol. 267, pp. 4281-4284 (1992).
Murthy et al, Clin. Chem., vol. 38, pp. 1307-1310 (1992).
Kobayashi et al, Transplantation Proceedings, vol. 25, pp. 655-657 (1993).
Kobayashi Masakazu
Niwa Mineo
Ohtsuka Kazuyuki
Tanaka Hirokazu
Fujisawa Pharmaceutical Co. Ltd.
Hutzell Paula K.
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