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Updated: May 21, 2026
Optimized Production and Analysis of Recombinant Protein-Filled Vesicles from E. coli
Published on: June 30, 2023
Protocol for recombinant expression in E. coli and purification of iBoost vaccine proteins using immobilized metal
Elisabeth J M Huijbers1, Judy R van Beijnum2, Joost Koetsier2
1Angiogenesis Laboratory, Department of Medical Oncology, Amsterdam UMC, Cancer Center Amsterdam, 1081 HV Amsterdam, the Netherlands; CimCure BV, 1066 CX Amsterdam, the Netherlands.
The iBoost vaccine technology enables the induction of a humoral immune response against self- and low-immunogenic antigens through fusion of the antigen to an immunogenic bacterial carrier. Here, we present a protocol for recombinant expression in E. coli and purification of iBoost vaccine proteins using immobilized metal affinity chromatography (IMAC). In addition, we describe procedures to evaluate antigen-specific immune responses in mice. This protocol can be readily adapted for the generation and testing of vaccines against a broad range of antigens. For complete details on the use and execution of this protocol, please refer to Huijbers et al., Van Beijnum et al., and Van Loon et al.1,2,3,4.
The iBoost vaccine technology enables the induction of a humoral immune response against self- and low-immunogenic antigens through fusion of the antigen to an immunogenic bacterial carrier. Here, we present a protocol for recombinant expression in E. coli and purification of iBoost vaccine proteins using immobilized metal affinity chromatography (IMAC). In addition, we describe procedures to evaluate antigen-specific immune responses in mice. This protocol can be readily adapted for the generation and testing of vaccines against a broad range of antigens. For complete details on the use and execution of this protocol, please refer to Huijbers et al., Van Beijnum et al., and Van Loon et al.1,2,3,4.
