Peptide Phage Display: Molecular Principles and Biomedical Applications

Zambrano-Mila, Marlon S.; Blacio, Karen Elizabeth Sánchez; Vispo, Nelson Santiago

doi:10.1007/s43441-019-00059-5

Cited by 27 publications

(20 citation statements)

References 67 publications

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“…Since George Smith first established the phage display system in 1985, numerous bacteriophage species have been engineered for phage display, including M13, f1, T4, and phage λ ( Zambrano-Mila et al, 2020 ). Phage display technology is beneficial to several biological fields due to its powerful mechanism for production of high copy numbers of peptides or proteins, the analysis of protein interactions ( Pande et al, 2010 ), the isolation of functional compounds, and the study of antigen–antibody binding ( Hamzeh-Mivehroud et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Biological Characterization and Evolution of Bacteriophage T7-△holin During the Serial Passage Process

Bao

Hong

et al. 2021

Front. Microbiol.

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Bacteriophage T7 gene 17.5 coding for the only known holin is one of the components of its lysis system, but the holin activity in T7 is more complex than a single gene, and evidence points to the existence of additional T7 genes with holin activity. In this study, a T7 phage with a gene 17.5 deletion (T7-△holin) was rescued and its biological characteristics and effect on cell lysis were determined. Furthermore, the genomic evolution of mutant phage T7-△holin during serial passage was assessed by whole-genome sequencing analysis. It was observed that deletion of gene 17.5 from phage T7 delays lysis time and enlarges the phage burst size; however, this biological characteristic recovered to normal lysis levels during serial passage. Scanning electron microscopy showed that the two opposite ends of E. coli BL21 cells swell post-T7-△holin infection rather than drilling holes on cell membrane when compared with T7 wild-type infection. No visible progeny phage particle accumulation was observed inside the E. coli BL21 cells by transmission electron microscopy. Following serial passage of T7-△holin from the 1st to 20th generations, the mRNA levels of gene 3.5 and gene 19.5 were upregulated and several mutation sites were discovered, especially two missense mutations in gene 19.5, which indicate a potential contribution to the evolution of the T7-△holin. Although the burst size of T7-△holin increased, high titer cultivation of T7-△holin was not achieved by optimizing the culture process. Accordingly, these results suggest that gene 19.5 is a potential lysis-related component that needs to be studied further and that the T7-△holin strain with its gene 17.5 deletion is not adequate to establish the high-titer phage cultivation process.

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Section: Introductionmentioning

confidence: 99%

Biological Characterization and Evolution of Bacteriophage T7-△holin During the Serial Passage Process

Bao

Hong

et al. 2021

Front. Microbiol.

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“…This approach also allows a thorough characterization of the antigenic epitopes by screening a series of overlapping peptides covering the entire primary sequence of the antigen by high-throughput approaches, [40]. Alternatively, phage display libraries of sufficiently high complexity may be used [41]. However, several disadvantages can arise when using peptide-based approaches; (i) three dimensional or conformational epitopes consisting of distant residues are not represented by linear peptides, (ii) posttranslational modifications are difficult to be recapitulated by synthetic peptides, and (iii) signal strength may be considerably lower with a single short peptide than with the full-length protein, in view that the autoimmune response in a given individual is usually polyclonal and several antigenic determinants are simultaneously recognized by different immunoglobulins.…”

Section: Discussionmentioning

confidence: 99%

Autoimmunity to the Follicle-Stimulating Hormone Receptor (FSHR) and Luteinizing Hormone Receptor (LHR) in Polycystic Ovarian Syndrome

Schniewind

Sattler

Haudum

et al. 2021

IJMS

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Hyperandrogenemia and ovulatory dysfunction are hallmarks of polycystic ovary syndrome (PCOS), pointing to a deranged hypothalamus-pituitary-ovarian (HPO) axis. An autoimmune etiology of PCOS is suspected in a subset of patients due to the relatively high concordance of PCOS with common autoimmune diseases. For this reason, we tested the hypothesis that natural autoantibodies (aAb) to the follicle-stimulating hormone receptor (FSHR) or luteinizing hormone receptor (LHR) are prevalent in PCOS. To this end, new luminometric assays for quantifying aAb to the FSHR (FSHR-aAb) or LHR (LHR-aAb) were developed using full-length recombinant human receptors as fusion proteins with luciferase as reporter. Prevalence of FSHR-aAb and LHR-aAb was determined in serum samples from healthy controls and PCOS patients. Steroid hormone profiles were compared between patients with and without FSHR-aAb or LHR-aAb. Signal linearity and detection ranges were characterized and both methods passed basic performance quality checks. The analysis revealed a relatively low prevalence, with 4 out of 430 samples positive for FSHR-aAb in the control versus 11 out of 550 samples in the PCOS group, i.e., 0.9% versus 2.0%, respectively. Similarly, there were only 5 samples positive for LHR-aAb in the control versus 2 samples in the PCOS group, i.e., 1.2% versus 0.4%, respectively. Samples positive for FSHR-aAb displayed steroid hormones in the typical range of PCOS patients, whereas the two samples positive for LHR-aAb showed relatively elevated free testosterone in relation to total testosterone concentrations with unclear significance. We conclude that the FSHR and LHR constitute potential autoantigens in human subjects. However, the prevalence of specific autoantibodies to these receptors is relatively low, both in control subjects and in women with PCOS. It is therefore unlikely that autoimmunity to the LHR or FSHR constitutes a frequent cause of hyperandrogenemia or ovulatory dysfunction in PCOS.

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“…Phage display technology involves the expression of sequences of interest inserted within a gene encoding a viral capsid protein, and a modified target peptide is subsequently displayed on the viral capsid of the phage (22). Phage display technology has developed tremendously and changed several fields, such as oncology, cell biology, immunology, pharmacology, and drug discovery (23).…”

Section: Discussionmentioning

confidence: 99%

Construction and Identification of New Molecular Markers of Triple-Negative Breast Cancer Stem Cells

et al. 2021

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Objective: We screened the TNBC stem cells using phage display (PD) and acquired the specific binding clones; and then the positive phage DNAs were amplified and extracted, synthesized with specific polypeptides, and labeled with fluorescein isothiocyanate (FITC). Finally, we identified the specificity of the polypeptides in vitro and in vivo.Methods: Human breast cancer cell line MDA-MB-231 and human mammary gland cell line hs578bst were chosen in our study, and MDA-MB-231 breast cancer stem cells (BCSCs) were cultured and identified by flow cytometry. The phage peptide library was screened using MDA-MB-231 BCSCs, the positive phage clones were identified by ELISA, and the DNA of the positive phages was extracted and sent to a biotechnology company for sequencing. According to the sequencing results, a specific polypeptide was synthesized and labeled with FITC. In the end, the specificity of a polypeptide to BCSCs was identified in vivo and in vitro.Results: The MDA-MB-231 BCSCs were cultured and enriched with the “serum and serum-free alternate” method. The BCSCs were found to have characteristics of CD44+/CD24−/low epithelial surface antigen (ESA) and ALDH+ with flow cytometry. The phage was enriched to 200-fold after three rounds of screening for MDA-MB-231 BCSCs. The positive phages were sequenced; then a polypeptide named M58 was synthesized according to sequencing results. Polypeptide M58 has a specific affinity to MDA-MB-231 BCSCs in vivo and in vitro.Conclusion: Specific polypeptides binding to MDA-MB-231 BCSCs were screened out by PD screening method, which laid a theoretical foundation for the targeted therapy and further research of BCSCs.

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Peptide Phage Display: Molecular Principles and Biomedical Applications

Cited by 27 publications

References 67 publications

Biological Characterization and Evolution of Bacteriophage T7-△holin During the Serial Passage Process

Biological Characterization and Evolution of Bacteriophage T7-△holin During the Serial Passage Process

Autoimmunity to the Follicle-Stimulating Hormone Receptor (FSHR) and Luteinizing Hormone Receptor (LHR) in Polycystic Ovarian Syndrome

Construction and Identification of New Molecular Markers of Triple-Negative Breast Cancer Stem Cells

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