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Updated: Dec 31, 2025
Antigenic Liposomes for Generation of Disease-specific Antibodies
Published on: October 25, 2018
[Preparation of mouse polyclonal antibodies against lipoprotein SPD_1609 in Streptococcus pneumoniae]
Xiaoyan Yang1, Liyuan He1, Fachang Wei1
1Zhuhai Key Laboratory of Basic and Applied Research in Chinese Medicine, Department of Bioengineering, Zhuhai Campus of Zunyi Medical University, Zhuhai 519041, China.
Abstract:
Objective To prepare mouse polyclonal antibodies against lipoprotein SPD_1609 in Streptococcus pneumoniae. Methods Firstly, the spd_1609 gene in Streptococcus pneumoniae (S. pneumoniae) D39 strain was amplified and ligated into the prokaryotic expression vector of pGEX-4T-1 to generate recombinant plasmid pGEX-4T-1609. Secondly, the fusion protein of GST-1609 was expressed in Escherichia coli BL21 (DE3), and purified using GST affinity chromatography. After cleaving the GST-tag of the fusion protein, the SPD_1609 protein was obtained and purified using GST affinity chromatography. Finally, BALB/c mice were immunized with the purified SPD_1609 protein to produce polyclonal antibodies. The titer and specificity of polyclonal antibodies were detected using ELISA and Western blotting. Results In this study, the recombinant vector of pGEX-4T-1609 was constructed, and the SPD_1609 protein (Mr is approximately 35 000) was expressed and purified with a higher 95% purity. The ELISA results suggested that the SPD_1609 protein could induce specific immune responses in mice, and the titer of antibody in the immunized mice was 1:40 960. Western blotting indicated that the polyclonal antibody could specifically recognize the SPD_1609 protein expressed in prokaryotic expression vector and in S. pneumoniae. Conclusion The mouse polyclonal antibody with good specificity against SPD_1609 protein has been successfully obtained.
Insights
Researchers successfully developed mouse polyclonal antibodies targeting the SPD_1609 lipoprotein in Streptococcus pneumoniae. These antibodies demonstrate high specificity and titer, crucial for potential diagnostic or therapeutic applications against pneumococcal infections.
Area of Science:
- Microbiology
- Immunology
- Molecular Biology
Background:
- Streptococcus pneumoniae is a significant human pathogen.
- SPD_1609 is a lipoprotein found in S. pneumoniae.
- Developing specific antibodies is crucial for understanding and combating bacterial infections.
Purpose of the Study:
- To generate mouse polyclonal antibodies against the SPD_1609 lipoprotein.
- To characterize the specificity and titer of the produced antibodies.
Main Methods:
- Gene amplification and cloning of spd_1609 into a prokaryotic expression vector.
- Expression and purification of the SPD_1609 protein using GST affinity chromatography.
- Immunization of BALB/c mice with purified SPD_1609 protein.
- Antibody characterization via ELISA and Western blotting.
Main Results:
- A recombinant plasmid pGEX-4T-1609 was successfully constructed.
- The SPD_1609 protein was expressed and purified to >95% purity.
- ELISA confirmed a high antibody titer of 1:40,960.
- Western blotting demonstrated specific recognition of SPD_1609 by the antibodies.
Conclusions:
- Mouse polyclonal antibodies with high specificity against SPD_1609 protein were successfully produced.
- These antibodies can recognize SPD_1609 in both recombinant forms and within S. pneumoniae.
- The developed antibodies are valuable tools for further research into S. pneumoniae pathogenesis.
