中文版 English
 
新闻动态
公司新闻
最新促销
行业新闻
技术视频
联系我们

英国Ludger生物科技有限公司中国办事处

电 话:021-60398329

邮 箱:lily.wang@ludger.com & cindy.li@ludger.com

地 址:上海静安区南京西路1266号

当前位置:英国Ludger生物科技有限公司中国办事处>>最新促销>>唾液酸(Sialic Acid Analysis )分析相关产品

唾液酸(Sialic Acid Analysis )分析相关产品

Ludger Sialic Acid Analysis Guide I: NeuAc and NeuGc Quantitation

Sialic acid analysis is a regulatory requirement laid out in the ICH Q6B guidelines for characterisation of biopharmaceuticals. In particular, determination of relative levels of human and non-human type sialic acids (NeuAc/NANA and NeuGc/NGNA respectively) is important since some patients have high levels of anti-NeuGc antibodies which could lead to neutralization and rapid clearing of NeuGc-containing biopharmaceuticals (Nguyen et al, 2005).

The significance of NeuAc and NeuGc when analysing glycoproteins means that it is often considered a critical quality attribute (CQA) for biotherapeutics.

A widely used method for determining the ratio of NeuAc to NeuGc is as follows:

• Release of sialic acid residues from the glycoprotein by mild acid hydrolysis.

• Fluorescent labeling of released sialic acids with1,2-diamino-4,5-methylenoxybenzene (DMB).

• Relative quantitative analysis of DMB-labelled sialic acids by HPLC or UHPLC.

The LudgerTag™ DMB kit (Cat No. LT-KDMB-A1) provides all that is required to release sialic acids from glycoproteins and label them with DMB. The kit contains reagents and materials for up to 22 glycoprotein samples (50-100 μg of glycoprotein per sample). Each labeling should work with approximately 10 pmol up to 2.5 nmol sialic acids per sample.

Included in the kit are a sialic acid reference panel (containing Neu5Ac, Neu5Gc, Neu5,7Ac2, Neu5,Gc9Ac, Neu5, 9Ac2 and Neu 5,7, (8), 9Ac3Gc) and both NeuAc and NeuGc quantitative standards. These three standards can be labelled with DMB alongside the released sialic acid samples.

Preparation of serial dilutions of the NeuAc and NeuGc quantitative standards enables quantitative analysis. Information on how to do this is explained in the LudgerTag™ DMB kit guide. The NeuAc and NeuGc quantitative standards are also available to purchase separately if required.

For positive controls, we recommend using fetuin glycoprotein which contains both NeuAc and NeuGc sialic acids. We also recommend a purified glycopeptide standard, the first in a range of Ludger BioQuant™ quantitative standards, as a positive control. This glycopeptide standard (Cat No. BQ-GPEP-A2G2S2-10U) is a complex biantennary N-linked glycan terminating in two N-acetylneuraminic acids.

Using this standard will enable you to check the efficiency of glycan release, labeling and recovery and will give you confidence in the accuracy of your sialic acid measurements. Ludger also sells a standard which can be used for identification of Neu5,9Ac2. Water can be used as a negative control.

ReferencesNguyen DH, Tangvoranuntakul P, and Varki A. (2005) Effects of Natural Human Antibodies against a Nonhuman Sialic Acid That Metabolically Incorporates into Activated and Malignant Immune Cells. J Immunol 175:228-236

Figure 1: DMB-labeled sialic acid reference panel profiled on an LS-uR2 UHPLC column (Cat No. LS-UR2-2.1x100). The reference panel can be used as a system suitability standard prior to the sample run in order to check separation of glycans.

Sialic acid profiling related products

商品型号

英文品名

中文品名

包装

LT-KDMB-A1

DMB Sialic Acid Labeling Kit

DMB 唾液酸释放标记试剂盒

1Reaction Set

LS-R1-4.6x150

Ludger Sep R1 HPLC Column

R1 HPLC 色谱柱-唾液酸

4.6 x150 mm

LS-UR2-2.1x100

Ludger Sep UR2 UHPLC Column–sialic acids

UR2 UHPLC 色谱柱-唾液酸

2.1 x 100 mm

CM-NEU-AC-01

N-acetylneuraminic acid quantitative standard

N-乙酰神经氨酸定量标准品

1 nmol

CM-NEU-GC-01

N-glycolyneuraminic acid quantitative standard

N-羟乙酰神经氨酸定量标准品

1 nmol

CM-NEU5,9,AC-01

N-acetylneuraminic acid qualitative standard

N-乙酰神经氨酸定性标准品

1 nmol

BQ-GPEP-A2G2S2

A2G2S2 glycopeptide standard

A2G2S2 糖肽标准品(用于唾液酸释放和标记过程)

10 μg

GCP-FET-50U-X4

Fetuin Glycoprotein (4 x 50 ug)

胎球糖蛋白(4×50微克)(唾液酸化阳性对照)

4 x 50 μg

 

友情链接:   Ludger英国   |   谷歌   |   百度  

Copyright @ 2009-2022   英国Ludger生物科技有限公司中国办事处 沪ICP备20140708号 技术支持:溢尚网络
电 话:021-60398329 邮 箱:lily.wang@ludger.com & cindy.li@ludger.com 地 址:上海静安区南京西路1266号