Application of Sodium Dodecyl Sulfate-Capillary Gel Electrophoresis to the Characterization of Ricin A-Chain Immunotoxins

作者：Na Dong Hee; Park Eun Ji; Kim Myung Sun; Lee Hye Suk; Lee Kang Choon^*

来源：Chromatographia, 2012, 75(11-12): 679-683.

DOI：10.1007/s10337-012-2243-3

• Sodium dodecyl sulfate-capillary gel electrophoresis
• Immunotoxins
• Monoclonal antibody
• Ricin

This study was undertaken to use sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) for the characterization of immunotoxins constructed by cross-linking ricin A-chain (RTA) to monoclonal antibody (MAb). SDS-CGE could separate mono- and di-RTA-conjugated MAbs from unreacted MAb, as identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Resolution of RTA-MAb conjugates was improved by decreasing the running voltage from 15 to 5 kV. SDS-CGE was also useful for characterizing aggregates of RTA or deglycosylated RTA (dgRTA) that was formed during immunotoxin preparation. Of the RTA isomers with different carbohydrate contents, RTA(1), which has low carbohydrate content, was found to be more susceptible to aggregation than RTA(2). In the SDS-CGE of dgRTA, both covalently and non-covalently linked aggregates were observed under reducing and non-reducing conditions. The findings of this study suggest that SDS-CGE can be used to optimize methods of preparing effective immunotoxins and for checking degradants and aggregates.

• 出版日期2012-6