Antibody compositions with altered fab sialylation

A sialylation and antibody technology, applied in the fields of antibody, drug combination, organic chemistry, etc.

Active Publication Date: 2019-09-20
CSL BEHRING AG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although many antibodies are currently in use, there remains a need to provide alternative and / or improved antibody preparations with more beneficial effectiveness in clinical applications

Method used

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  • Antibody compositions with altered fab sialylation
  • Antibody compositions with altered fab sialylation
  • Antibody compositions with altered fab sialylation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0042] The term "antibody" as used throughout the present invention encompasses, for example, polyclonal or monoclonal antibodies. The term "antibody" also includes derivatives or fragments thereof that retain antigen-binding specificity. Thus, embodiments such as chimeric (human constant regions, non-human variable domains), single chain and humanized (human antibodies with non-human CDRs) antibodies are also included, as well as antibody fragments, e.g., particularly , Fab or Fab' fragment. Antibody fragments or derivatives also include Fd, F(ab') 2 , Fv or scFv fragments; see, e.g., Harlow and Lane "Antibodies, A Laboratory Manual", Cold Spring Harbor Laboratory Press, 1988 and Harlow and Lane "Using Antibodies: A Laboratory Manual", Cold Spring Harbor Laboratory Press, 1999.

[0043] The term "Fab region" as used throughout the present invention refers to the region of an antibody consisting of one constant and one variable domain from each of the heavy and light chains ...

Embodiment 1

[0122] [Example 1: Fractionation of IVIG using 2,6-sialic acid-specific Sambucus nigra lectin (SNA)]

[0123] 1 g IVIG in 100 ml tris-buffered saline (TBS), 0.1 mM CaCl at pH 9-10 2 Recirculate for 1 h on a 100 ml Sambucus nigra lectin (SNA) column. Flow through with collection 100ml TBS / CaCl 2washing. After washing with 2 x 200ml of TBS, the fraction bound to the SNA column (+SA IVIG) was eluted with 200ml of 0.5M lactose in 0.2M acetic acid. Run a 2nd flow-through fraction on the column to obtain -SA IVIG ( figure 1 ).

[0124] Total sialic acid content in IgG was monitored by SDS-PAGE (non-reducing conditions) and lectin blot ( figure 1 ). The resulting IVIG fractions were separated by SDS-PAGE using Nupage 10% BisTris gel. Gels were stained with Coomassie and blotted to nitrocellulose. Blots were probed with biotin-SNA and AP-streptavidin and visualized with chromogenic substrates. exist figure 1 In D, heavy and light chains were separated by SDS-PAGE under reduc...

Embodiment 2

[0128] [Example 2: Glycan analysis]

[0129] To study the glycosylation profile of IVIG before and after isolation, identify and characterize tryptic glycopeptides derived from the Fc region of typical IVIG preparations, and characterize the total N-glycan population of IVIG by glycan release and analysis.

[0130] 【method】

[0131] [(a) Determination of IgG1 and IgG2 / 3 Fc Glycan Characterization by Peptide Localization]

[0132] IVIG samples were analyzed by liquid chromatography-mass spectrometry (LC-MS) after trypsinization to determine glycan profiles specific to IgG1 and IgG2 / 3 Fc regions. Glycopeptides that monitor the IgG2 Fc region are also found in the IgG3 Fc region, hence they are referred to as IgG2 / 3. Based on the known relative abundance of IgG subclasses in serum, most of the signal seen for IgG2 / 3 peptides was expected to originate from IgG2 molecules. Denaturation, reduction (heating with guanidinium-HCl and DTT) and alkylation (with iodoacetamide) followed...

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Abstract

The present invention relates to an antibody population enriched from an antibody preparation, wherein the enriched antibody population has an altered amount of sialylation in the Fab region of the antibody compared to the antibody preparation prior to enrichment. Furthermore, the present invention relates to a method of enriching an antibody population from an antibody preparation, wherein the enriched antibody population has an altered amount of sialylation in the Fab region of the antibody compared to the antibody preparation prior to enrichment. The present invention also relates to the antibody population of the invention for use in medicine, pharmaceutical compositions comprising the antibody population of the invention and the antibody population of the invention in the prevention and / or treatment of atherosclerosis, cancer and infections such as bacteria, viruses or fungi Use in infection.

Description

[0001] 【Technical field】 [0002] The invention relates to populations of antibodies with altered sialylation in the Fab region, methods of making them, and uses thereof. [0003] 【Background technique】 [0004] Immunoglobulins, also known as antibodies, are the main secretory products of the immune system. They are generally formed from basic structural units - each having 2 heavy chains and 2 light chains - to form monomers with one such unit, dimers with 2 units, pentamers with 5 units, Or a hexamer with 6 units. Antibodies play a prominent role in innate immunity. In a natural immune response against a pathogen, a complex is formed between the pathogen and the antibody. These immune complexes activate a wide range of effector functions, thereby leading to the killing, removal and destruction of pathogens. Antibodies can also react with the body's own antigens which can lead to autoimmune diseases and contribute to chronic inflammatory symptoms. Antibodies can have anti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/00C07K1/22A61K39/395A61K39/00
CPCA61K2039/505A61P7/04A61P9/10A61P17/00A61P21/00A61P25/00A61P25/16A61P25/28A61P29/00A61P31/04A61P31/10A61P31/12A61P35/00A61P37/02C07K1/22C07K16/00C07K16/081C07K16/088C07K16/1027C07K16/1081C07K16/1282C07K2317/21C07K2317/41C07K2317/54C07K2317/55
Inventor F·卡瑟曼M·乔迪S·梅斯舍尔K·埃尔康
Owner CSL BEHRING AG
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