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High-throughput sequencing enhanced phage display identifies peptides that bind mycobacteria
Nqobile A C Ngubane1, Lionel Gresh, Thomas R Ioerger
1Emerging Health Technologies Platform, Council for Scientific and Industrial Research, Biosciences Unit, Pretoria, Gauteng, South Africa ; KwaZulu-Natal Research Institute for Tuberculosis and Human Immunodeficiency Virus, Nelson R. Mandela School of Medicine, University of KwaZulu-Natal, Durban, South Africa.
Phage display technology identified novel peptides that bind to Mycobacterium tuberculosis. This approach is powerful for discovering non-antigenic biomarkers for tuberculosis and other bacterial infections.
Area of Science:
- Microbiology
- Biotechnology
- Immunology
Background:
- Bacterial cell wall components are established infection biomarkers detectable by antibodies.
- Mycobacterium tuberculosis (M. tb) may possess non-antigenic surface molecules, complicating biomarker discovery using traditional antibody-based methods.
Purpose of the Study:
- To employ phage display technology to identify peptides that bind to mycobacteria.
- To compare the efficacy of random clone picking versus high-throughput sequencing for identifying enriched clones.
Main Methods:
- Phage display technology was utilized to screen for peptides binding to mycobacteria.
- Clone identification was performed using both random picking and high-throughput sequencing (Illumina).
- Surface Plasmon Resonance (SPR) was used to validate peptide-target interactions.
Main Results:
- High-throughput sequencing identified the CPLHARLPC peptide as the most enriched clone.
- The CPLHARLPC peptide demonstrated superior binding to mycobacteria compared to clones selected by random picking.
- SPR confirmed that synthesized CPLHARLPC peptide binds to a 15 KDa peptide from M. tb H37Rv lysates.
Conclusions:
- Phage display combined with high-throughput sequencing is an effective strategy for identifying novel peptide biomarkers.
- This method can uncover potential non-antigenic biomarkers for tuberculosis (TB) and other bacterial infections.
- The identified CPLHARLPC peptide shows promise for further investigation in TB diagnostics.
