Creating a Compact Human Antibody Library Using Phage Display
Author Information
Author(s): Pansri Potjamas, Jaruseranee Nanthnit, Rangnoi Kuntalee, Kristensen Peter, Yamabhai Montarop
Primary Institution: Suranaree University of Technology
Hypothesis
Can a highly efficient method for constructing a human scFv library be developed using a diverse repertoire from non-immunized donors?
Conclusion
The study successfully demonstrates a method to create a compact and efficient phage display library with high diversity from non-immunized human donors.
Supporting Evidence
- The library consisted of 1.5 × 10^8 individual clones derived from 140 non-immunized human donors.
- Antibodies were successfully isolated against a variety of antigens, including complex ones like snake venom and cancer cell surface antigens.
- The method used fewer oligonucleotide primers and steps compared to previous library construction methods.
Takeaway
Scientists found a way to make a library of tiny proteins that can stick to different things, using blood from healthy people instead of animals.
Methodology
The library was constructed using a simple method involving PCR amplification of variable gene segments from B-lymphocytes of 140 non-immunized donors, followed by cloning into a phagemid vector.
Potential Biases
Potential bias in the selection of donors and the specific antigens used for testing may affect the generalizability of the findings.
Limitations
The study may not account for the full diversity of the human antibody repertoire as it is based on a specific population of non-immunized donors.
Participant Demographics
Participants were healthy, non-immunized donors aged between 17-50 from Nakhon Ratchasima, Thailand.
Digital Object Identifier (DOI)
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