Characterization of monoclonal antibodies on the Agilent 1260 Infinity Bio-inert Quaternary LC by Size Exclusion Chromatography using the Agilent BioSEC columns
نویسندگان
چکیده
The Agilent 1260 Bio-inert Quaternary LC System and Agilent BioSEC columns were used to investigate the purity and stability of two monoclonal antibodies (mAb): AntiFLAG and BL05. The high sensitivity of the Agilent 1260 Infinity Diode Array Detector was used to characterize these antibodies at low concentrations. The bio-inert LC system in combination with BioSEC columns resulted in the development of reliable and rugged methods amenable for routine size exclusion chromatography of therapeutic mAbs and proteins. For both the mAbs, superior separation of variants from the main antibody peak was achieved and compared to the results obtained with a competitive column. Authors Srividya Kailasam, Jochen Strassner, Faizy Ahmed and Martin Vollmer Agilent Technologies Bangalore, India and Waldbronn, Germany Application Note
منابع مشابه
pH Gradient Elution for Improved Separation of Monoclonal Antibody Charge Variants
Ion exchange is a very useful technique to separate protein mixtures under mild conditions, most commonly by applying a shallow gradient of increasing salt concentration as eluent. pH gradient elution is used much less frequently, except in specialist applications such as chromatofocusing, and can require complex buffer systems. However, today’s modern quaternary HPLC systems are ideally suited...
متن کاملDevelopment and partial validation of a SEC method for high-resolution separation and quantifi cation of monoclonal antibodies
Size exclusion chromatography (SEC) is an important technique for the characterization of high molecular substances such as proteins or polymers. It is the standard method to determine and quantify aggregation levels for protein-based pharmaceuticals and is generally required for regulatory approval. This Application Note describes a simple and sensitive method for the quantitative analysis of ...
متن کاملOptimizing Protein Separations with Agilent Weak Cation-Exchange Columns
Columns containing weak cation-exchange stationary phases offer a degree of versatility when used for separation of positively charged proteins. Of particular interest, is the difference in selectivity that can be obtained by adjusting the pH of the mobile phase eluent. We explore the parameters of interest and how these can be changed to optimize the resolution of the compounds of interest usi...
متن کاملOptimizing protein separations with cation exchange chromatography using Agilent Buffer Advisor Protein separation with the Agilent 1260 Infinity Bio‐inert Quaternary LC System
This Technical Overview shows that the Agilent Buffer Advisor software in com‐ bination with the Agilent 1260 Infinity Bio‐inert Quaternary LC System is an ideal solution for automated protein separation by ionic strength gradients. Usually, pH scouting using premixed two‐component gradients is time‐consuming and work‐ intensive. Dynamically mixed four‐component gradients calculated by the Buff...
متن کاملSemi - Preparative Purifi cation and Characterization of Lysozyme Modifi ed with Poly Ethylene Glycol ( PEG ) The Agilent 1260
This Application Note used hen egg white lysozyme as a model protein for purifi cation and biophysical characterization of PEGylated proteins. PEGylation to the N-terminus of lysozyme in a site-specifi c manner was achieved using PEG-propionaldehyde derivatives with a molecular weight of Mw 20,000. A semi-preparative RP-HPLC method for the purifi cation of PEGylated lysozyme was developed using...
متن کامل