Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1990-11-21
1993-01-12
Chan, Y. Christina
Chemistry: molecular biology and microbiology
Spore forming or isolating process
43524026, 5303881, 5303882, 5303887, 53038873, 53038875, 53038885, 935103, C12N 520, C07K 1528, C12P 2108
Patent
active
051790190
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a new hybridoma cell line and to the monoclonal antibody produced by this hybridoma cell line This monoclonal antibody reacts with a novel human leucocyte surface membrane antigen with wide distribution within the haemopoietic system.
BACKGROUND OF THE INVENTION
Since it was shown by Kohler & Milstein (Nature Vol. 256, 495-497, 1975) that it was possible to fuse mouse myeloma cells with spleen cells from immunized mice and thereby product a continuous cell line which produces a homogeneous (monoclonal) antibody, extensive attention has been focused on the production of these hybrid cell lines (hybridomas) and the monoclonal antibodies (Mabs) produced.
The development of hybridoma technology has led to a dramatically improved understanding of the antigenic molecules on the surface of human leucocytes, and over the past decade, many murine monoclonal antibodies reacting with haemopoietic cell surface antigens have been described. Following the three International Workshops on Human Leucocyte Differentiation Antigen, the majority of these antibodies have been grouped into Clusters of Differentiation (CD), and have been shown to react with restricted differentiation-lineage or maturation stage membrane antigens on lymphoid or bone marrow-derived cells (Interim Report of the 3rd Workshop Nomenclature Committee (1987) in A.J. McMichael, Ed. Leucocyte Typing III. White cell differentiation antigen. Oxford University Press, PP 949-950).
A few antibodies have been shown to have broader haemopoietic cellular reactivity. The best characterised are the antibodies fitting into CD 45 or CD 45R, which identify a family of antigens with molecular weights around 200 Kilodaltons expressed on virtually all human leucocytes (Pizzolo et al., Cancer, 1980, 46, 2640-2647; Dalchau et al., 1980, European Journal of Immunology, 10, 737-744). Other unclustered Mabs with non-lineage reactivity include PHM-1, CAMPATH-1, HuLyM3, which appear to identify separate and unique antigens (Becker et al., 1981, Pathology, 13, 669-680; Hale et al., 1983, Blood, 62, 873-882; Vaughan et al., Transplantation, 36, 446-450).
SUMMARY OF THE INVENTION
In a first aspect, the present invention consists in a mouse monoclonal antibody of class IgG.sub.1 produced by a hybridoma formed by a fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with the T cell line HSB-2 and T-CLL cells, the monoclonal antibody being characterised in that it reacts with a leucocyte surface membrane antigen of a relative molecular weight of approximately 40-50 Kilodaltons which is expressed on over 90% of normal human peripheral blood mononuclear cells and granulocytes, but not on normal platelets or erythrocytes.
It is preferred that the monoclonal antibody is further characterised in that it:
In a second aspect the present invention consists in an IgG.sub.1 monoclonal antibody-producing hybridoma cell line formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with the T cell line HSB-2 and T-CLL cells, the monoclonal antibody produced being characterised in that it reacts with a leucocyte surface membrane antigen of a relative molecular weight of approximately 40-50 Kilodaltons which is expressed on over 90% of normal human peripheral blood mononuclear cells and granulocytes, but not on platelets or erythrocytes.
It is preferred that the monoclonal antibody produced is further characterised in that it:
The monoclonal antibody of the first aspect of the present invention has been designated WM-65, and the hybridoma cell line of the second aspect of the present invention has been designated F56-1D5. This hybridoma cell line was deposited with the European Collection of Animal Cell Cultures (ECACC), PHLS Centre for Applied Microbiology and Research, Porton Down, Salisbury, Wilts., United Kingdom on Mar. 30, 1989 and was accorded accession number 89033001.
The disclosure of this deposit is herein incorporated by way of cross-refe
REFERENCES:
LeBien et al., The Journal of Immunology, vol. 125, No. 5, pp. 2208-2214 (1980).
Atkinson Michael K.
Bradstock Kenneth F.
Henniker Anthony J.
Biomedical Systems Limited
Chan Y. Christina
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