Neuraminic Acid Analysis

Neuraminic Acids (sialic acids) are most often found at the terminal position of glycans, and therefore are a likely point of contact for glycan-ligand interactions. Neuraminic acid analysis is a regulatory requirement laid out in the ICH Q6B guidelines for the characterization of biopharmaceuticals.

Biopharmaceutical sialylation refers to the type and distribution of sialic acids in the glycans of therapeutic glycoproteins. They are important for the stability and 3D conformation of glycoproteins and are involved in many biologic interactions. Neuraminic acids have an important functional impact: sialyation of the N-glycans on IgG increases anti-inflammatory activity; O-acetylated neuraminc acids can change ligand interactions and affect degradation) O-acetylated sialic acids including Neu5,9Ac2 are found on EPO; and the presence of neuraminic acids also increases the serum half-life of glycoproteins by preventing uptake by the asialoglycoprotein receptor located on liver cells.

Furthermore, the sialylation pattern can be a very useful measure of product consistency during manufacturing. Advances in our understanding of these issues have led the FDA, EMEA and other regulators to tighten their rules on glycoprofiling throughout the drug life cycle.

A diverse range of neuraminic acids are found in nature, but the two main types of neuraminic acid residues found in biopharmaceuticals produced in mammalian expression systems are N-acetyl-neuraminic acid (Neu5Ac or NANA) and N-glycolylneuraminic acid (Neu5Gc or NAGA). These usually occur as terminal structures attached to galactose (Gal) residues at the non-reducing terminii of both N- and O-linked glycans. Humans cannot synthesize Neu5Gc and its presence on a drug can lead to immune reactions such as chronic inflammation. Anit-Neu5Gc antibodies have been detected in normal human sera and can neutralize any Neu5Gc containing biopharmaceuticals, thereby lowering the drug’s efficacy. It is important to be aware that the choice of cell line can greatly influence the type of neuraminic acids present on a biopharmaceutical, for example a large portion of the sialic acids on mouse IgG are often Neu5Gc.

Controlling the ratio of NeuAc to NeuGc is critical for biomanufacturers. It is therefore imperative for drug safety and efficacy to monitor both the level and types of neuraminc acids during all stages of the product life cycle as QC batch-to-batch consistency.

Ludger Quantitative Sialic Acid Analysis Application Note

DMB Sialic Acid Labeling Kit

The LudgerTag DMB Sialic Acid Labeling Kit has been developed for the qualitative analysis of sialic acids
The kit contains reagents for the release of sialic acids from glycoproteins. Released sialic acids are conjugated with DMB dye by an amination-cyclisation reaction.

The kit contains reagents and materials for up to 22 samples including the sialic acid reference panel, and the N-acetyl neuraminic acid and N-glycolyl neuraminic acid standards.

View product documentation as PDF
View Certificate of Analysis as PDF

Part Number: LT-KDMB-A1

uR2 2.1x100mm UHPLC Column

The LudgerSep uR2 2.1x100mm column has been developed for UHPLC analysis of sialic acids labeled with DMB using the LudgerTag DMB Sialic Acid Labeling Kit.

Dimensions: 2.1mm x 100mm

View product documentation as PDF
View Certificate of Analysis as PDF

Part Number: LS-UR2-2.1x100

Sialic Acid Quantitation Kit

The SialiQuant Sialic Acid Quantitation Kit contains all the reagents to quickly and accurately quantitate sialic acids, including N-acetylneuraminic acid (NANA), N-glycolylneuraminic acid (NGNA).

View product documentation as PDF

Part Number: KE-SIALIQ

Sialic Acid Aldolase

Converts sialic acid to N-acetylmannosamine & pyruvate. Sialic acids can then be quantitated by a coupled reaction with NADH and lactic dehydrogenase

View product documentation:
Specsheet    CoA   MSDS

Part Number: E-ALD01

CMP-Sialic Acid Synthetase

This enzyme catalyzes the synthesis of CMP sialic acid from CTP & sialic acid.
The resulting molecule can then be covalently attached to an oligosaccharide by sialic acid transferase.

View product documentation as PDF

Part Number: E-CMP01

Sialidase Au

recombinant from A. ureafaciens

View product documentation:
Specsheet    CoA    MSDS

Part Number - Amount of Enzyme
E-S001       - 60 µLs¹
E-S001-20   - 20 µLs¹
E-S001-200 - 200 µls²
   ¹ includes buffer
   ² includes enzyme only

$180$1,344

Sialidase Cp

Sialidase Cp cleaves all non-reducing terminal non-branched α(2-3) and α(2-6) sialic acid residues from complex carbohydrates and glycoproteins.

recombinant from C. perfringens

View product documentation:
Specsheet    CoA    MSDS

Part Number - Amount of Enzyme
E-S005       - 60 µLs¹
E-S005-20   - 20 µLs¹
E-S005-200 - 200 µls²
   ¹ includes buffer
   ² includes enzyme only

$180$1,344

Sialidase Sp

Sialidase Sp cleaves the non-reducing terminal α(2-3) unbranched sialic acid residues from complex carbohydrates and glycoproteins.

recombinant from Streptococcus pneumoniae in E. Coli

View product documentation:
Specsheet   CoA    MSDS

Part Number - Amount of Enzyme
E-S007       - 60 µLs¹
E-S007-20   - 20 µLs¹
E-S007-200 - 200 µls²
   ¹ includes buffer
   ² includes enzyme only

$180$1,344

NANA standard – Quantitative

The N-acetylneuraminic acid (NANA) standard is a quantitative standard of NIST-F and USP traceable Neu5Ac monosaccharide.

View product documentation as PDF
Download Non-hazardous MSDS

Part numbers:
    CM-NEU-AC-01   1 nmols
    CM-NEUAC-100  100 nmols

NGNA Standard – Quantitative

The N-glycoylneuraminic acid (NGNA) standard is a quantitative standard of NIST-F and USP traceable Neu5Gc monosaccharide.

View product documentation as PDF
Download Non-hazardous MSDS

Part numbers:
    CM-NEU-GC-01   1 nmols
    CM-NEUGC-100  100 nmols

Neu5,9Ac2

5-N-acetyl-9-O-acetyl neuraminic acid standard (Neu5,9Ac2)

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Download Non-hazardous MSDS

~1 nmol (of which >750 pmol is Neu5,9Ac2)

Part Number: CM-NEU5,9AC2-01

Sialic Acid Reference Panel

The Sialic Acid Reference panel is a mixture of different sialic acids forms (as detailed on chromatogram below) sourced from bovine submaxillary mucin.

It includes the following sialic acids: Neu5Ac; Neu5Gc; Neu5,7Ac2; Neu5,Gc9Ac; Neu5,9Ac2; Neu5,7,(8),9Ac3Gc.

Amount: approx. 1.25 nmols

View product documentation as PDF
Download Non-hazardous MSDS

Part Number: CM-SRP-01