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  • Title: [Conjugation of Plasmodium falciparum Pfs25 to Pseudomonas aeruginosa ExoProtein A with different chemical linkers].
    Author: Qian F.
    Journal: Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi; 2011 Aug; 29(4):254-7. PubMed ID: 21972597.
    Abstract:
    OBJECTIVE: To conjugate Plasmodium falciparum Pfs25 to recombinant Pseudomonas aeruginosa Exo-Protein A (rEPA) and test the effect of conjugation using different chemical linkers. METHODS: The Pfs25 was thiolated by NAHT (DL-N-acetylhomocysteine thiolactone), whereas the carrier protein rEPA was modified by Sulfo-EMCS, EMCH, SBAP and Sulfo-SIAB respectively. The Pfs25-rEPA conjugates were formed by incubating the thiolated Pfs25 with modified rEPA under certain conditions. Coomassie blue stained SDS-PAGE was performed to examine the results of the conjugation. RESULTS: The Pfs25 was successfully conjugated to the carrier protein rEPA by the linkers used in this study. For the addition of a chemical group onto the protein, the reaction between the groups of primary amine and NHS (N-hydroxysuccinimide) ester was more efficient than the reaction between the groups of carboxyl and hydrazide with EDC as a crosslinker, for the formation of a conjugate, the reaction between the groups of maleimide and free sulfhydryl was more efficient than the reaction between the groups of haloacetyl and free sulfhydryl. CONCLUSION: The Pfs25 can be conjugated to the rEPA by the chemical linkers with different conjugation efficiency and coupling proteins.
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