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

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Abstract

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, RTA1, which has low carbohydrate content, was found to be more susceptible to aggregation than RTA2. 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.

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Acknowledgments

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0025345), and by the Ministry of Knowledge Economy (MKE) and Korea Institute for Advancement in Technology (KIAT) through the Workforce Development Program in Strategic Technology.

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Authors and Affiliations

  1. College of Pharmacy, Kyungsung University, Busan, 608-736, Republic of Korea

    Dong Hee Na, Eun Ji Park & Myung Sun Kim

  2. College of Pharmacy, The Catholic University of Korea, Bucheon, 420-743, Republic of Korea

    Hye Suk Lee

  3. Drug Targeting Laboratory, College of Pharmacy, SungKyunKwan University, Suwon, 440-746, Republic of Korea

    Kang Choon Lee

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  1. Dong Hee Na
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  2. Eun Ji Park
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  3. Myung Sun Kim
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Correspondence to Kang Choon Lee.

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Na, D.H., Park, E.J., Kim, M.S. et al. Application of Sodium Dodecyl Sulfate-Capillary Gel Electrophoresis to the Characterization of Ricin A-Chain Immunotoxins. Chromatographia 75, 679–683 (2012). https://doi.org/10.1007/s10337-012-2243-3

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  • DOI: https://doi.org/10.1007/s10337-012-2243-3

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