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ANTIBODIES TO CD1d

An antibody and antigen technology, applied in the direction of antibodies, anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve problems such as unmet medical needs

Inactive Publication Date: 2014-11-12
TEVA PHARMA AUSTRALIA PTY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is a significant unmet medical need in the treatment of patients with severe forms of asthma, such as corticosteroid-refractory asthma

Method used

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  • ANTIBODIES TO CD1d
  • ANTIBODIES TO CD1d
  • ANTIBODIES TO CD1d

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0220] Example 1-Generation of anti-CD1d antibodies

[0221] Phage display

[0222] FAbs that bind to CD1d / β2M in humans and cynomolgus monkeys were isolated from the phagemid library used for the first experiment.

[0223] Anti-CD1d / β2MFAbs were isolated from the phage display library during the two panning "activities" (i.e., isolated phage display tests with different reagents or panning conditions). The general test procedure follows the method outlined by Marks et al. (Marks, J.D. & Bradbury, A., 2004, Methods Mol Biol, 248, 161-66).

[0224] Each phage display event includes three rounds of panning. For each round, by mixing 1:1 with blocking buffer (5% skim milk in phosphate buffered saline, pH 7.4) and incubating for 1 hour at room temperature, the ~1×10 13 The phage particles are blocked. The blocked phage library was then incubated with 100 microliters of streptavidin-conjugated Dynabead (Invitrogen) for 45 minutes to pre-exclude the streptavidin binding agent, and the str...

Embodiment 2

[0237] Example 2-Confirmation of IgG binding to CD1d

[0238] The human-cynomolgus CD1d-reactive FAb was converted into IgG4 format, expressed and purified as described in the general method. The purified antibodies were tested for binding to human and cyno CD1d by ELISA and SPR using the modified version of the assay described in Example 1. In short, for the ELISA assay, a Maxisorp ELISA plate (Nunc) was coated at 1 μg / ml with the appropriate antigen. The plate was then washed and the purified IgG sample was added to the individual wells of the ELISA plate. The IgG was allowed to bind to the captured CD1d / β2M at room temperature for one hour, and then washed three times with PBS-T and three times with PBS. The bound IgG was detected using HRP-conjugated antibody against human Fc (Sigma). The detection antibody was incubated for 30 minutes at room temperature. The plate was washed to remove unbound antibody, and the assay signal was visualized by incubating with 50 microliter...

Embodiment 3

[0242] Example 3-Cell-based CD1d tetramer inhibition titer determination

[0243] Generate stable NKTCR expressing cell line

[0244] In order to develop a cell-based assay that characterizes the biological potency of anti-CD1d antibodies, a stable cell line expressing the NKT cell receptor (NKTCR) is required. The cell line J.RT3-T3.5 (ATCC:TIB-153) was selected to generate a stable NKT cell receptor expressing cell line. J. RT3-T3.5 is derived from the E6-1 clone of Jurkat lacking the β chain of the T cell antigen receptor (ATCC: TIB152). The cell does not express CD3 or the T cell receptor αβ heterodimer on the surface. J.RT3-T3.5 cells were electroporated with two vectors, one containing the α chain of J3N.5NKTCR (SEQ ID NO: 21), and the other containing the β chain of J3N.5NKTCR (SEQ ID NO: 22) (Brigl, M. et al., 2006 JImmunol 176:3625-34.). This NKT cell receptor is reactive to the glycolipid antigen α-GalCer. These vectors encoding the α and β chains of the NKTCR also e...

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Abstract

The present invention provides isolated antibodies or antigen binding portions thereof which bind to CD1d. These antibodies and antigen binding portions thereof have application in treatment of conditions involving NKT cell effector function.

Description

[0001] submit data [0002] This application is related to the Australian Patent Application No. 2011904190 filed on October 14, 2011 and the U.S. Patent Application No. 61 / 547,307 filed on October 14, 2011 and claims priority. The entire contents of each of these applications are incorporated herein by reference. This article. Invention field [0003] The present invention relates to antibodies that bind to CD1d and inhibit CD1d-mediated biological functions (for example, activation of CD1d restricted T cells, natural killer T (NKT) cells). Background of the invention [0004] The bibliographic details of the publications mentioned by the author in this manual are collected at the end of the manual in alphabetical order. [0005] The mention of any prior art in this specification is not and should not be regarded as an acknowledgement or any form of suggestion that prior art forms part of common general knowledge in any country. [0006] CD1d is an essential counter-receptor for trig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K16/28A61P11/06A61P17/06A61P1/16A61P9/10A61P43/00C12N15/13C12N5/10
CPCC07K2317/55C07K16/2833C07K2317/34C07K2317/565C07K2317/76C07K16/2803A61K2039/505C07K2317/52C07K2317/33C07K2317/92A61P1/04A61P1/16A61P11/00A61P11/06A61P17/06A61P43/00A61P9/10
Inventor J.K.纳姆比亚尔L.D.波顿A.克拉克A.J.波D.塔姆瓦基斯G.科普斯达斯A.G.多伊勒M.波拉德H.穆斯塔发
Owner TEVA PHARMA AUSTRALIA PTY LTD
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