Jayachandran R

Jayachandran R

United States Pharmacopeia-India Pvt. Ltd., India

Title: An alternate approach for sialic acid estimation in erythropoietin by HPAEC-PAD method


• Jayachandran is currently working with USP as a senior manager and responsible for overseeing the development of documentary and reference standards for various biotherapeutic products such as Mabs, Proteins and Peptides • He has around 12 years of experience in the area of analytical method development, validation and Physico-chemical characterization. • Exploring high-end analytical techniques like Q-TOF, protein sequencing and gained in-depth understanding about the application of such tools in protein characterization. • Experience in setting up of laboratory and quality management system • Academically, holding a Post Graduate degree in Pharmacy with specialization in biotechnology. Worked with reputed biotech organizations like INTAS, NEKTAR and SANOFI.


High-pH anion exchange chromatography method coupled with pulsed amperometric detection (HPAEC-PAD) was developed as an alternative to calorimetric and flourometric based methods for estimation of sialic acid in Erythropoietin (EPO). N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) are most commonly determined sialic acids in glycoproteins. Using this method, these two sialic acids were successfully determined without derivitization in EPO samples. Samples were treated with neuraminidase enzyme isolated from Arthobacter ureafaciens followed by addition of internal standard 2-keto -3-deoxy-D-glycero-D-galacto-2 nonulosonic acid (KDN). Analysis was performed on Carbopac PA20 column with disposable gold electrode on PTFE using acetate buffer system in NaOH. Commercially available Neu5Ac and Neu5Gc were used as calibration standards. The responses of standards were found to be linear from 50 to 250 pmol for Neu5Ac and 2 to 10 pmol for Neu5Gc with r2 ≥ 0.99 respectively. This method was further evaluated for specificity, linearity, accuracy and precision to determine its suitability for routine analysis of sialic acid in EPO samples. Hence, this method can be used as an alternate method to colorimetric methods for quantifying the Sialic acid content.

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