IEF simulation is an effective tool to investigate the transport phenomena and separation performance as well as to design IEF microchip. However, multidimensional IEF simulations are computationally intensive as one has to solve a large number of mass conservation equations for ampholytes to simulate a realistic case. In this study, a parallel scheme for a 2D IEF simulation is developed to red...

A robust and highly reproducible capillary isoelectric focusing (cIEF) method for the evaluation of charge heterogeneity of monoclonal antibody (mAb) pharmaceutical which contains covalently bound antitumor compounds was developed using a combination of commercially available dimethylpolysiloxane-coated capillary and carrier ampholyte. In order to optimize major analytical parameters for robust...

PROTEINS are generally least hydrated and least soluble at their respective isoelectric points. In this connection the process of electrofocusing of proteins (Svenson, 1962; Vesterberg & Svenson, 1966) is relevant. In this method several proteins are electrophoretically propelled-through an electrophoretically established pH gradient-until they reach the particular pH regions which equal their ...

Background and Aims: Influenza virus nucleoprotein (NP) has the capacity to be used as subunit vaccine, but little is known about the impact of different cultures on its structure. In the present study we aimed to evaluate and compare the Isoelectric focusing (IEF) property of extracted viral nucleoproteins derived from Madin Darby canine kidney (MDCK) cell line and embryonated chicken eggs (EC...

This chapter provides guidance and procedures used for characterization of biotechnology-derived articles by amino acid analysis. This chapter is harmonized with the corresponding chapter in JP and EP. Portions of the chapter that are not harmonized with the other two pharmacopeias are marked by the symbol ♦. The footnote below is in the USP but is not in the EP or JP. Other characterization te...

The pharmaceutical industry and most particularly the biopharmaceutical are demanding advancements in analytical techniques. field of proteomics peptide mapping needs sensitive analysis sophisticated instrumentation, which hyphenated techniques to be explored. In addition effective detection characterization, Due efficient selective separation, predominant focus has been drawn capillary electro...

Numerical simulations are presented for ampholyte-based isoelectric focusing in 2D microgeometries. In this study, model proteins are focused in the presence of 25 biprotic ampholytes under an applied electric field. Each protein is considered as a simple polypeptide having ten charge states, while the biprotic ampholytes are selected to generate a shallow pH range of 6 to 9. Straight and contr...

The separation of ampholytic components according to isoelectric point has played an important role in isolating, reducing complexity and improving peptide and protein detection. This brief review outlines the basics of isoelectric focusing, including a summary of the historical achievements and considerations in experimental design. Derivative methodologies of isoelectric focusing are also dis...