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Human antibody molecules for IL-13

A human antibody, antibody technology, applied in the direction of antibodies, anti-cytokine/lymphokine/interferon immunoglobulin, anti-inflammatory agents, etc.

Inactive Publication Date: 2015-10-21
MEDIMMUNE LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, IL-13 inhibitors may prevent disease progression by inhibiting the proliferation of malignant Reed-Sternberg cells

Method used

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  • Human antibody molecules for IL-13
  • Human antibody molecules for IL-13
  • Human antibody molecules for IL-13

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0270] Isolation of anti-IL-13 scFv

[0271] Repertoire of scFv antibodies

[0272] Screening was performed using a large single-chain Fv (scFv) human antibody library derived from splenic lymphocytes from 20 donors and cloned into a phagemid vector [66].

[0273] Screening of scFv

[0274] IL-13-recognizing scFvs were isolated from the phage-display library by performing a series of repeated rounds of selection on recombinant bacterial-derived human or murine IL-13 (Peprotech) essentially as described in [67]. Briefly, after incubation with the library, immobilized antigen pre-coupled to paramagnetic beads and bound phage were recovered by magnetic separation, while unbound phage were washed away. Bound phage were then rescued by the method described by Vaughan et al. [67] and by repeating the above screening process. Use different solid-phase surfaces and capture methods in different screening cycles to reduce non-specific binding. Antigens were either covalently coupled...

Embodiment 2

[0279] Neutralizing potency of anti-IL-13 in IL-13-dependent TF-1 cell proliferation assay

[0280] Neutralizing potency of purified scFv preparations against human and murine IL-13 bioactivity was assessed by TF-1 cell proliferation assay. Purified scFv preparations were prepared as described in Example 3 of WO01 / 66754. Protein concentrations of purified scFv preparations were determined using the BCA method (Pierce). TF-1 is a human myeloblastic cell line established from patients with erythroleukemia [68]. The survival and proliferation of the TF-1 cell line is factor-dependent. In this regard, TF-1 cells are responsive to human or murine IL-13 [69] and can be maintained in media containing human GM-CSF (4 ng / ml, R&D Systems). Inhibition of IL-13-dependent proliferation was determined by measuring the reduction of deuterium-labeled thymidine incorporated into newly synthesized DNA of dividing cells.

[0281] TF-1 Cell Assay Protocol

[0282] TF-1 cells were obtained fr...

Embodiment 3

[0286] Neutralizing potency of the lead clone obtained by optimizing parental clone heavy chain CDR3 in IL-13-dependent TF-1 cell proliferation assay

[0287] Osbourn et al. [70] demonstrated that directed mutagenesis of residues within the CDR3 of the heavy chain significantly increased antibody affinity. The scFv repertoire was screened as described in Example 1, wherein residues within the heavy chain CDR3 of BAK278D6 (SEQ ID NO: 6), BAK167A11 (SEQ ID NO: 57) had been randomized by mutagenesis. Unique clones among the products obtained from this screen were identified by DNA sequencing, and their neutralizing potency as scFv in the TF-1 cell proliferation assay was evaluated as described in Example 2.

[0288] result

[0289] Both pedigrees showed significant increases in potency. The most effective clones in the BAK167A11 lineage are BAK615E3, BAK612B5 and BAK582F7, and the ICs of the three as scFv against 25ng / ml human IL-13 in the TF-1 cell proliferation assay 50 3nM ...

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Abstract

There are described recombinant VH and VL domains of human antibodies that bind IL-13. A series of proteins characterised by three CDR in each VH / VL are described. Preferably the molecules are single chain Fv or antibody molecules. Nucleic acids expressing the VH and VL domains are selected from phage display libraries. Neutralization potency of the resulting scFv's was tested in TF proliferation assays. The fragments and antibodies showed efficacy for IL-13 binding in murine models of pulmonary inflammation, the primate (cynomolgus) model of dermal allergy, IgE release from B cells and histamine potentiation in Ca2+ signalling in smooth muscle. Mice transgenic for human IL-13 antibodies and epitope mapping studies are also described. It is envisaged that the anti 11-13 reagents may be used to treat asthma, atopic dermatitis, allergic rhinitis, fibrosis, IBS, or Hodgkin's lymphoma.

Description

[0001] This application is a divisional application of the national application number 200480026557.7 (PCT / GB2004 / 003059) filed on July 15, 2004 entitled "Human Antibody Molecules Against IL-13". technical field [0002] The present invention relates to specific binding members, in particular human anti-IL-13 antibody molecules, especially those human antibody molecules that neutralize the activity of IL-13. The invention further relates to the use of anti-IL-13 in the diagnosis or treatment of IL-13 related disorders including asthma, atopic dermatitis, allergic rhinitis, fibrosis, inflammatory bowel disease and Hodgkin's lymphoma Antibody Molecule Method. Background technique [0003] Preferred embodiments of the invention employ the antibody VH and / or VL domains of the antibody molecule referred to herein as BAK502G9 as well as other antibody molecules in the BAK502G9 lineage and BAK278D6 lineage as defined herein. A further preferred embodiment utilizes the BAK278D6 lin...

Claims

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Application Information

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IPC IPC(8): C07K16/24C12N15/13A61K39/395A61P11/06A61P17/00A61P11/02A61P37/08A61P1/00A61P29/00A61P35/00A61P11/00A61P19/04A61P37/00
CPCC07K16/244A61K2039/505C07K2317/21C07K2317/565C07K2317/622C07K2317/73C07K2317/76Y10S514/826Y10S514/883A61P1/00A61P11/00A61P11/02A61P11/06A61P17/00A61P19/04A61P27/16A61P29/00A61P35/00A61P37/00A61P37/06A61P37/08A61P43/00C07K16/24A61K39/395C12N15/11C07K2317/92
Inventor P.D.蒙克L.耶尔穆图斯R.R.明特C.P.索罗克
Owner MEDIMMUNE LTD
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