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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
Cost-effective HRMA pre-sequence typing of clone libraries; application to phage display selection
Barry A Pepers1, Menno H Schut, Rolf Ham Vossen
1Center for Human and Clinical Genetics, Leiden University Medical Center, Albinusdreef 2, Leiden, The Netherlands. b.a.pepers@lumc.nl
BMC Biotechnology
|May 26, 2009
Summary
High-resolution melt curve analysis (HRMA) offers a fast, cost-effective method for analyzing clonal variation without DNA sequencing. This technique efficiently identifies identical clones, reducing overall research costs.
Area of Science:
- Molecular Biology
- Biotechnology
Background:
- Comparing numerous clones is essential for methods like phage display and mutagenesis.
- High-resolution melt curve analysis (HRMA) presents a novel, cost-effective solution for clonal variation studies.
Purpose of the Study:
- To evaluate HRMA as a tool for rapid clonal characterization.
- To demonstrate HRMA's utility in reducing the need for extensive DNA sequencing.
Main Methods:
- High-resolution melt curve analysis (HRMA) was employed to analyze clonal variations.
- Results were compared with ELISA and DNA fingerprinting techniques.
Main Results:
- HRMA results demonstrated strong concordance with ELISA and DNA fingerprinting.
- DNA sequencing confirmed that HRMA effectively clustered identical clones.
Conclusions:
- HRMA enables simultaneous analysis of up to 384 samples in approximately 30 minutes.
- Automated software facilitates clone clustering, significantly reducing sequencing costs and providing insights into phage display antibody selection success.
