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Improved method to raise polyclonal antibody using enhanced green fluorescent protein transgenic mice
Jianke Ren1, Long Wang, Guoxiang Liu
1Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Acta Biochimica Et Biophysica Sinica
|February 1, 2008
Summary
Using transgenic mice expressing enhanced green fluorescent protein (EGFP) can improve antibody production. This method generates higher titers of desired antibodies and reduces unwanted immune reactions for better antibody preparation.
Area of Science:
- Immunology
- Biotechnology
- Protein Engineering
Background:
- Recombinant fusion proteins are common antigens for antibody production.
- Anticarier antibodies in antiserum reduce desired antibody yield and cause non-specific immune reactions.
- Carrier protein transgenic animals offer a potential solution to these issues.
Purpose of the Study:
- To investigate the efficacy of using carrier protein transgenic animals for raising high-titer polyclonal antibodies.
- To validate the hypothesis that transgenic animals can mitigate anticarier antibody responses.
Main Methods:
- Production of enhanced green fluorescent protein (EGFP) transgenic mice.
- Immunization of EGFP transgenic mice and wild-type mice with a His6HA tag-EGFP fusion protein.
- Analysis of antibody titers against the His6HA tag and EGFP in the resulting antiserum.
Main Results:
- Antiserum from EGFP transgenic mice exhibited a higher antibody titer against the His6HA tag compared to wild-type mice.
- Antiserum from EGFP transgenic mice showed a lower antibody titer against EGFP compared to wild-type mice.
- Demonstrated a reduction in antibodies against the carrier protein (EGFP).
Conclusions:
- EGFP transgenic mice provide an improved method for raising high-titer antipeptide polyclonal antibodies.
- This approach minimizes anticarier antibody production and non-specific immune responses.
- The method holds potential for broader applications in antibody preparation and biotechnology.
