Antibodies against human il-12
AssigneeHoffman-LaRoche Inc.
InventorsMaurice Kent Gately, David Howard Presky
Life of this patent
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Abstract
The present invention relates to novel p75 heterodimer specific anti-human IL-12 antibodies that are characterized by a higher potency and greater efficacy in neutralizing human IL-12 bioactivity than known heterodimer specific IL-12 monoclonal antibodies. The heterodimer specific antibodies recognize one or more epitopes of the human IL-12 p75 heterodimer, but do not bind to the p40 subunit alone. The heterodimer specific IL-12 antibodies neutralize rhesus monkey IL-12 bioactivity with a potency similar to their potency for neutralizing human IL-12 bioactivity making them useful IL-12 antagonists for in vivo studies in the rhesus monkey.
Claims31
A method for producing an antibody that immunologically reacts with the human IL-12 p75 heterodimer which consists of a p35 subunit and a p40 subunit, comprising the steps of:
- (a) immunizing a mammal deficient in a gene encoding an IL-12 p35 subunit or an IL-12 p40 subunit with the human IL-12 p75 heterodimer to produce antibodies;
- (b) obtaining antibodies from the immunized mammal;
- (c) screening said antibodies for their ability to bind an epitope presented by the p75 heterodimer to obtain said binding antibody.
A method for producing a monoclonal antibody that immunologically reacts with the human IL-12 p75 heterodimer which consists of a p35 subunit and a p40 subunit, comprising the steps of:
- (a) immunizing a mammal deficient in a gene encoding an IL-12 p35 subunit or an IL-12 p40 subunit with the human IL-12 p75 heterodimer to produce antibodies;
- (b) harvesting antibody producing cells from the immunized mammal;
- (c) forming a monoclonal antibody producing hybridoma from said cells and obtaining said monoclonal antibody;
- (d) screening said monoclonal antibody produced by said hybridoma for the ability to bind to an epitope presented by the p75 heterodimer to obtain said binding monoclonal antibody.
A monoclonal antibody to human IL-12 which consists of a p35 subunit and a p40 subunit forming a p75 heterodimer, wherein said monoclonal antibody,
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12, but is not immunologically reactive with any epitope presented by said p40 subunit;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.03 μg/ml or less when measured in an IL-12-stimulated PHA-activated human lymphoblast proliferation assay wherein the concentration of said human IL-12 is 0.25 ng/ml.
A monoclonal antibody to human IL-12 which consists of a p35 subunit and a p40 subunit forming a p75 heterodimer, wherein said monoclonal antibody,
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12, but is not immunologically reactive with any epitope presented by said p40 subunit;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.02 μg/ml or less when measured in an IL-12 stimulated IFN-γ production assay by PHA-activated human lymphoblasts, wherein the concentration of said human IL-12 is 0.25 ng/ml.
A monoclonal antibody to human IL-12 p75 heterodimer, said p75 heterodimer comprising a p40 subunit and a p35 subunit, wherein said monoclonal antibody:
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.03 μg/ml or less when measured in an IL-12-stimulated PHA-activated human lymphoblast proliferation assay wherein the concentration of said human IL-12 is 0.25 ng/ml.
A monoclonal antibody to human IL-12 p75 heterodimer, said p75 heterodimer comprising a p40 subunit and a p35 subunit, wherein said monoclonal antibody:
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.02 μg/ml or less when measured in an IL-12 stimulated IFN-γ production assay by PHA-activated human lymphoblasts, wherein the concentration of said human IL-12 is 0.25 ng/ml.
A monoclonal antibody to human IL-12 p75 heterodimer, said p75 heterodimer comprising a p40 subunit and a p35 subunit, wherein said monoclonal antibody:
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.03 μg/ml or less when measured in an IL-12-stimulated PHA-activated human lymphoblast proliferation assay wherein the concentration of said human IL-12 is 0.25 ng/ml;
- (c) is produced from a hybridoma cell line obtained by immunizing a mouse deficient in a gene encoding a p40 subunit or a p35 subunit of IL-12 with human IL-12 p75 heterodimer.
A monoclonal antibody to human IL-12 p75 heterodimer, said p75 heterodimer comprising a p40 subunit and a p35 subunit, wherein said monoclonal antibody:
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12;
- and
- (b) wherein the 50% inhibitory concentration (IC50) of said antibody for the bioactivity of human IL-12 is 0.02 μg/ml or less when measured in an IL-12 stimulated IFN-γ production assay by PHA-activated human lymphoblasts, wherein the concentration of said human IL-12 is 0.25 ng/ml;
- (c) is produced from a hybridoma cell line obtained by immunizing a mouse deficient in a gene encoding a p40 subunit or a p35 subunit of IL-12 with human IL-12 heterodimer.
A method for treating an autoimmune disease in an individual in need thereof, comprising administering to said individual a composition comprising a monoclonal antibody formulated in a pharmaceutically acceptable carrier, said monoclonal antibody binding to human IL-12 p75 heterodimer, said p75 heterodimer comprising a p40 subunit and a p35 subunit, wherein said monoclonal antibody:
- (a) immunologically reacts with an epitope presented by the p75 heterodimer of human IL-12;
- and
- (b1) neutralizes at least 90% of the bioactivity of human IL-12 as measured in an IL-12 bioactivity assay in PHA-activated human lymphoblasts by inhibiting IL-12 stimulated proliferation of PHA-activated human lymphoblasts, using an antibody concentration of 0.5 μg/ml and a human IL-12 concentration of 0.25 ng/ml;
- or
- (b2) neutralizes at least 90% of the bioactivity of human IL-12 as measured in an IL-12 bioactivity assay in PHA-activated human lymphoblasts by inhibiting IL-12 stimulated IFN-γ production by PHA-activated human lymphoblasts, using an antibody concentration of 0.5 μg/ml and a human IL-12 concentration of 0.25 ng/ml.
Dependencies are inferred from the claim text.