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C5 antigens and uses thereof

An antigen and antibody technology, applied in the direction of antibodies, peptide sources, instruments, etc., can solve problems such as cell death

Inactive Publication Date: 2010-03-24
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

C5b functions as a nucleation site for the assembly of C5b-9 (C5b, C6, C7, C8, and C9), also known as the terminal complement complex or the membrane attack complex that penetrates the cell membrane to form pores ( MAC), which may contribute to pro-inflammatory cell activation at sublytic concentrations but cause cell death at lytic concentrations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0260] In most cases, antibodies raised against human C5 did not show binding to mouse C5 due to the low conservation of the C5 protein sequence between mouse and human. Thus, chimeric C5 proteins (comprising human and mouse protein sequences) that retain activity in functional assays can be used to detect epitopes against human anti-C5-antibodies.

[0261] Antibodies can be mapped to epitopes on the human alpha or beta chain using DNA constructs expressing human alpha chain / mouse beta chain, or mouse alpha chain / human beta chain. DNA for the chimeric human / mouse C5 construct in plasmid form was obtained from GeneArt. The epitopes within the chain were finely mapped using a chimeric construct expressing a 100 amino acid segment of the mouse protein sequence that was grafted into the human C5 protein sequence in place of their respective human sequences. Each chimeric protein contains a string of histidines at its C-terminus for affinity purification.

[0262] The chimeric pr...

Embodiment 2

[0272] Example 2 Generation of human antibodies by phage display

[0273] To generate antibodies against C5, MorphoSys HuCAL Selection of phage display libraries. HuCAL GOLD is a Fab library based on the HuCAL concept in which all six CDRs are diversified and which uses CYSDISPLAY technology for ligation of Fab fragments to the phage surface (Knappik et al., 2000 J. Mol. Biol. 296:57-86; Krebs et al., 2001 J Immunol. Methods 254:67-84; Rauchenberger et al., 2003 J Biol Chem.278(40):38194-38205; WO 01 / 05950, , 2001).

[0274] Phagemid recovery (rescue), phage amplification and purification

[0275] The HuCAL GOLD library was amplified in 2xYT medium containing 34 μg / ml chloramphenicol and 1% glucose (2xYT-CG). use OD 600nm After infection with VCSM13 helper phage at 0.5 (30 minutes at 37°C, no shaking; shaking at 250 rpm for 30 minutes at 37°C), the cells were pelleted by centrifugation (4120 g; 5 minutes; 4°C), and resuspended in 2xYT / 34 μg / ml chloramphenicol / 50 μg...

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Abstract

The present invention pertains to the use of a complement component C5 inhibitor in methods of treatment of ocular disorders and the use of a complement component C5 inhibitor in the manufacture of amedicament in the treatment of an ocular disorder.

Description

Background of the invention [0001] Macular degeneration is a medical condition primarily found in older adults in which the center of the lining of the eye, called the macula, suffers from thinning, shrinkage and in some cases bleeding. This can lead to a loss of central vision, including the inability to see fine details, read, or recognize faces. The pathology of neochoroidal vascularization is poorly understood, but factors such as inflammation, ischemia and local production of angiogenic factors are thought to be important. [0002] Genes for complement system proteins have been identified to be strongly associated with a person's risk of developing macular degeneration. The complement system is a key component of innate immunity against microbial infection and consists of a group of proteins normally present in an inactive state in serum. These proteins are organized in three activation pathways: classical pathway, lectin pathway and alternative pathway. Molecules on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/00
CPCG01N2800/16G01N33/6893G01N2333/4716A61P27/02A61P27/04C12N15/11C07K14/47A61K39/395G01N33/53
Inventor B·C·吉尔德M·T·基廷M·米利克D·米哈伊洛夫M·罗古斯卡I·斯普劳斯基K·赵
Owner NOVARTIS AG
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