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Transient Expression of Proteins by Hydrodynamic Gene Delivery in Mice
Published on: May 5, 2014
Immunochemical evidence that human apoB differs when expressed in rodent versus human cells
Xingyu Wang1, Vinita Chauhan, Anh T Nguyen
1Lipoprotein and Atherosclerosis Research Group and the Department of Pathology, University of Ottawa Heart Institute, Ottawa, Ontario, Canada.
Journal of Lipid Research
|February 4, 2003
Summary
Researchers developed new monoclonal antibodies (MAbs) to study apolipoprotein B-100 (apoB-100) conformation. These MAbs revealed differences in apoB-100 structure between human and mouse models, impacting lipoprotein research.
Area of Science:
- Biochemistry
- Immunology
- Molecular Biology
Background:
- Low-density lipoprotein (LDL) from human apolipoprotein B-100 (apoB-100) transgenic mice exhibits higher triglyceride content compared to normolipidemic subjects.
- Understanding the conformation of apoB-100 is crucial for lipoprotein research and disease association studies.
Purpose of the Study:
- To generate novel monoclonal antibody (MAb) probes for investigating apoB-100 conformation.
- To characterize the epitopes recognized by anti-apoB-100 MAbs in different lipoprotein contexts.
Main Methods:
- Generation of hybridomas from human apoB-100 transgenic mice immunized with human LDL.
- Screening and characterization of anti-apoB-100 MAbs using native and denatured lipoproteins from various sources (human plasma, HepG2 cells, transgenic mice, transfected rat cells).
- Immunological assays including Western blotting and epitope mapping.
Main Results:
- Four anti-apoB-100 specific MAbs were identified, with two (2E1 and 3D11) recognizing an epitope in the N-terminal region of apoB-100.
- These MAbs detected the epitope on native lipoproteins from human plasma and HepG2 cells but not on lipoproteins from HuBTg+/+ mice or transfected rat cells.
- While 3D11 reacted strongly with denatured apoB-100 from HuBTg+/+ mice, the lack of reactivity with native murine lipoproteins suggests conformational or post-translational differences.
Conclusions:
- Human apoB-100 expressed in rodent systems (hepatocytes or hepatoma cells) adopts a different conformation or undergoes distinct post-translational modifications compared to human systems.
- The generated MAbs (2E1 and 3D11) are valuable tools for studying apoB-100 conformation and can differentiate between human and rodent-expressed apoB-100.
