Glycosylation on a protein is an important post-translational modification that can affect its function, clearance and stability. The PA 800 plus Pharmaceutical Analysis System simplifies the complex process of profiling carbohydrates associated with glycoproteins. By providing specific and accurate quantitation of glycosylation levels, differences in glycoform quantities and distribution can be determined.

Oligosaccharide Analysis Simplified
Following endonuclease cleavage from the protein, oligosaccharides are specifically labeled with amino-pyrene-trisulfonic acid (APTS) by reductive amination. This analysis is performed directly from the glycoprotein hydrolysate. With APTS derivatization and LIF detection, each sugar yields the same detector response, so their relative quantities can be directly compared.

Oligosaccharide distribution associated with a protein yields a fingerprint that can be used in protein identification. The PA 800 plus provides easy, quantitative and robust determination of protein microheterogeneity, with a typical analysistime of less than 15 minutes.

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Key benefits to carbohydrate profiling using capillary electrophoresis include:
	▪	Quantitation of N-linked oligosaccharides*
	▪	High resolution separation methods capable of differentiating positional isomers
	▪	Validated methodology developed for routine use environments
^*APTS-labeled carbohydrate analysis using capillary electrophoresis is a patented technology (US patent #US5,569,366).

Additional Information:
	▪	Carbohydrate Labeling and Analysis Chemistries
	▪	PA 800 plus Carbohydrate Labeling and Analysis Guide
	▪	"CE Separation of N-Linked Oligosaccharides Released from Recombinant Monoclonal Antibody" (AIB A-1986A) - This bulletin describes the isolation, separation and profiling analysis of complex N-linked oligosaccharides by CE using the Carbohydrate Labeling and Analysis Assay.
	▪	"Separation of Fucosylated, non-Fucosylated, and Complex Carbohydrates Associated with Monoclonal Antibodies using Capillary Electrophoresis" (P-14278A) - CE was used for automated and quantitative analysis of N-linked oligosaccharides. CE Chemistry was optimized and new methods were developed to enable the separation and differentiation of fucosylated, afucosylated, sialylated and complex antennary oligosaccharides in a simple, reproducible assay environment.
	▪	Other examples of Glycoprotein Analysis: Oligosaccharide Profiling, Monosaccharide Analysis, Glycoform Analysis

Click here for more information on Carbohydrate Labeling and Analysis Chemistries.

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