Antibody composition with altered fab sialylation

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

Active Publication Date: 2012-07-11
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 composition with altered fab sialylation
  • Antibody composition with altered fab sialylation
  • Antibody composition with altered fab sialylation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0038] 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.

[0039] 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

[0118] [Example 1: Fractionation of IVIG using 2,6 sialic acid-specific Sambucus nigra lectin (SNA)]

[0119] 1 g IVIG in 100 ml tris-buffered saline (TBS), 0.1 mM CaCl, pH 9-10 2 Recirculate for 1 h on a 100 ml Sambucus nigra lectin (SNA) column. Flow-through with collection of 100ml TBS / CaCl 2washing. After washing with 2 x 200 ml of TBS, the fraction bound to the SNA column (+SA IVIG) was eluted with 200 ml of 0.5M lactose in 0.2M acetic acid. Run the 2nd pass-through fraction on the column to obtain -SA IVIG ( figure 1 ).

[0120] Total sialic acid content in IgG was monitored by SDS-PAGE (non-reducing conditions) and lectin blotting ( figure 1 ). The obtained IVIG fractions were separated by SDS-PAGE using Nupage 10% BisTris gel. The gel was Coomassie stained 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 ...

Embodiment 2

[0124] [Example 2: Glycan analysis]

[0125] To study the glycosylation profile of IVIG before and after isolation, the tryptic glycopeptides derived from the Fc region of typical IVIG preparations were identified and characterized, and the total N-glycan population of IVIG was characterized by glycan release and analysis.

[0126] 【method】

[0127] [(a) Determination of IgG1 and IgG2 / 3 Fc glycan characteristics by peptide localization]

[0128] IVIG samples were analyzed by liquid chromatography-mass spectrometry (LC-MS) after trypsin digestion to determine glycan signatures specific to the IgGl and IgG2 / 3 Fc regions. The glycopeptides that monitor the IgG2 Fc region are also found in the IgG3 Fc region, so they are called IgG2 / 3. Based on the known relative abundance of IgG subclasses in serum, most of the signal seen for IgG2 / 3 peptides is expected to originate from IgG2 molecules. Denaturation, reduction (heating with guanidinium-HCl and DTT) and alkylation (with iodoac...

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Abstract

The present invention relates to a population of antibodies enriched from an antibody preparation, wherein the enriched population of antibodies has an altered amount of sialylation in the Fab region of the antibodies as compared to the antibody preparation prior to enrichment. Furthermore, the present invention relates to a method of enriching a population of antibodies from an antibody preparation, wherein the enriched population of antibodies has an altered amount of sialylation in the Fab region of the antibodies as compared to the antibody preparation prior to enrichment.; The present invention also relates to the population of antibodies of the invention for use in medicine, a pharmaceutical composition comprising the populations of antibodies of the invention and the use of the population of antibodies of the invention in the prevention and / or treatment of atherosclerosis, cancer and infections such as bacterial, viral or fungal infections.

Description

【Technical field】 [0001] The invention relates to populations of antibodies with altered sialylation in the Fab region, methods of making them, and uses thereof. 【Background technique】 [0002] Immunoglobulins, also known as antibodies, are the major secretory products of the immune system. They are generally formed from basic building blocks - 2 heavy and 2 light chains each - to form monomers with one of said units, dimers with 2 units, pentamers with 5 units, or a hexamer with 6 units. Antibodies play a significant role in innate immunity. In the natural immune response against pathogens, complexes are formed between pathogens and antibodies. 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 self-antigens that can lead to autoimmune diseases and contribute to symptoms of chronic inflammation. Antibodies can have anti-inflammatory activity,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C07K1/22A61K39/395A61K39/00
CPCC07K16/1027C07K16/1081C07K16/1282C07K2317/21C07K2317/54C07K16/081C07K1/22C07K16/00C07K16/088A61K2039/505C07K2317/41C07K2317/55A61P7/04A61P9/10A61P17/00A61P21/00A61P25/00A61P25/16A61P25/28A61P29/00A61P31/04A61P31/10A61P31/12A61P35/00A61P37/02
Inventor F·卡瑟曼M·鲁格瑟戈S·梅斯舍尔K·埃尔康
Owner CSL BEHRING AG
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