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Method for producing active GcMAF

A technology of active and activating factors, which is applied in the field of preparation of active GcMAF, can solve problems such as complex conversion procedures, and achieve efficient and simple steps

Pending Publication Date: 2020-08-11
FOUND FOR BIOMEDICAL RES & INNOVATION
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  • Abstract
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  • Claims
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Problems solved by technology

[0003] On the other hand, if figure 1 As schematically shown, the conversion process from inactive VDBP to active GcMAF is complicated, and in its preparation method, generally, inactive VDBP is purified from serum or plasma, and sialidase, Method of removing sialic acid and galactose by treating with β-galactosidase to form active GcMAF in which only GalNAc remains
However, such a conventional method requires a plurality of complicated steps such as a step of purifying inactive VDBP from serum or plasma, and a step of enzymatically treating the obtained VDBP. Therefore, a simpler and easier preparation method is required.

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  • Method for producing active GcMAF
  • Method for producing active GcMAF
  • Method for producing active GcMAF

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[0029]

[0030] The method for producing active GcMAF of the present invention is characterized in that it includes the step of culturing the cells introduced with the VDBP expression vector in a serum-free medium, and does not require an enzyme treatment step for cleaving sugar chains of inactive VDBP.

[0031] The preparation method of the active GcMAF of the present invention is not to purify the Gc globulin from plasma and serum to prepare the active GcMAF, but to cultivate the cells introduced into the VDBP expression vector in a serum-free medium without the need for many A method for easily preparing active GcMAF in one step.

[0032] In the present invention, VDBP is vitamin D binding protein (VDBP; Vitamin D Binding Protein), also known as Gc globulin, Gc protein, including three subtypes (1f, 1s, 2) with different sugar chain structures. In any subtype of VDBP, there is a common structure in which O-glycoside and galactose are bonded to N-acetylgalactosamine which ...

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Abstract

The purpose of the present invention is to provide a method for producing active GcMAF more simply and at a high yield. The present invention is a method for producing active GcMAF, the method including a step for culturing, in serum-free medium, host cells having a VDBP expression vector introduced therein. This culture is preferably a suspension culture. In addition, this method for producing active GcMAF is also characterized by not requiring an enzyme treatment step for sugar chain cleavage.

Description

technical field [0001] The invention relates to a preparation method of active GcMAF. Background technique [0002] Vitamin D binding protein (VDBP; Vitamin D Binding Protein) is one of the glycoproteins synthesized in the liver and secreted into the blood. VDBP binds to vitamin D and functions as a transport carrier in the blood. In addition, it is also known to bind to G-actin leaked from damaged cells to inhibit actin polymers from narrowing and blocking the vascular lumen. The aforementioned VDBP is also called Gc globulin, and there are three subtypes (VDBP1f, VDBP1s, and VDBP2) that differ in some amino acids and differ in sugar chain structure. Among them, in VDBP1f (Gc1f), the 418th (420th) threonine is bound to an O-type sugar composed of a trisaccharide in which sialic acid and galactose are bound to N-acetylgalactosamine (GalNAc) chain. The VDBP bound with the O-type sugar chain composed of the trisaccharide is an inactive type that does not have the function ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12P21/02
CPCC12P21/02C07K2319/21C07K14/47C07K1/22
Inventor 锅岛阳一锅岛曜子安部千秋
Owner FOUND FOR BIOMEDICAL RES & INNOVATION
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