Identification of Hapten-Carrier Protein Conjugate by Three Electrophoresis Methods

LEI Hong-tao1, PANG Jie1,2, HE Li-ping3, YANG Jin-yi1, SUN Yuan-ming1, QIN Yi-lan1, LIANG Ming-biao1 (1Guangdong Key Laboratory of Food Quality and Safety/College of Food Science, South China Agricultural University, Guangzhou 510642; 2College of Food Science and Technology, Fujian Agriculture and Forestry University, Fuzhou 350002; 3Instrumental Analysis and Research Center of South China Agricultural University, Guangzhou 510642)

hapten-carrier protein bioconjugate; SDS-PAGE; nondenaturing agarose gel electrophoresis; capillary zone electrophoresis

ObjectiveTo establish a simple electrophoresis characterization method for hapten-carrier protein bioconjugate. Method SDS-PAGE, nondenaturing agarose gel electrophoresis and high performance capillary electrophoresis were employed and compared with spectrum method. Result The result showed that SDS-PAGE was not a sensitive candidate method to differentiate carrier protein from bioconjugate when both proteins have less molecular weight difference, but nondenaturing gel electrophoresis was a better alternative, which could discriminate the free carrier protein and bioconjugate protein with different net charge numbers. High performance capillary zone electrophoresis was another powerful approach for hapten-carrier protein bioconjugate characterization. The information obtained from three electrophoresis was in accordance with the one from ultraviolet- visible spectrum scanning. Conclusion Three electrophoresis methods had different advantages and disadvantages, cost-saving and simple nondenaturing electrophoresis seemed to have more availability in most laboratories.