Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage

Hasmoni, Siti Salwa; Yusoff, Khatijah; Tan, Wen Siang

doi:10.2323/jgam.51.125

Cited by 7 publications

(3 citation statements)

References 16 publications

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“…Currently, bacteriophage M13 has been used widely in phage display technology and many peptides and proteins have been displayed successfully on the surface of the particle [6]. The displayed peptides and proteins have been applied in various fields of study including protein-protein interactions [7][8][9], identification of mimotopes for the development of vaccines [10,11], investigation of immune response [12,13], evaluation of small molecules drug candidates [14,15], identifying of peptide ligands for a variety of receptor ligands [16], and development of diagnostic reagents [6,[17][18][19][20]. Recently, M13 has been studied for its uses in nanostructures and nanotechnology for developing nanowires, nanosized energy storage unit and biocomposite fibers [21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Purification of bacteriophage M13 by anion exchange chromatography

Monjezi

Tey

Sieo

et al. 2010

Journal of Chromatography B

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M13 is a non-lytic filamentous bacteriophage (phage). It has been used widely in phage display technology for displaying foreign peptides, and also for studying macromolecule structures and interactions. Traditionally, this phage has been purified by cesium chloride (CsCl) density gradient ultracentrifugation which is highly laborious and time consuming. In the present study, a simple, rapid and efficient method for the purification of M13 based on anion exchange chromatography was established. A pre-packed SepFast Super Q column connected to a fast protein liquid chromatography (FPLC) system was employed to capture released phages in clarified Escherichia coli fermented broth. An average yield of 74% was obtained from a packed bed mode elution using citrate buffer (pH 4), containing 1.5 M NaCl at 1 ml/min flow rate. The purification process was shortened substantially to less than 2 h from 18 h in the conventional ultracentrifugation method. SDS-PAGE revealed that the purity of particles was comparable to that of CsCl gradient density ultracentrifugation method. Plaque forming assay showed that the purified phages were still infectious.

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Section: Introductionmentioning

confidence: 99%

Purification of bacteriophage M13 by anion exchange chromatography

Monjezi

Tey

Sieo

et al. 2010

Journal of Chromatography B

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“…Finally, the absorbance at 405 nm was determined in a microtiter plate reader. In this study, the LOD of phage-ELISA assay was about 10 ng with 1.0 × 10 12 pfu/mL of the purified fusion phage [ 23 ].…”

Section: Fluorescent Signal Generated By Genetically Engineered Phagesmentioning

confidence: 99%

Advances in the Development of Phage-Based Probes for Detection of Bio-Species

et al. 2022

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Bacteriophages, abbreviated as “phages”, have been developed as emerging nanoprobes for the detection of a wide variety of biological species, such as biomarker molecules and pathogens. Nanosized phages can display a certain length of exogenous peptides of arbitrary sequence or single-chain variable fragments (scFv) of antibodies that specifically bind to the targets of interest, such as animal cells, bacteria, viruses, and protein molecules. Metal nanoparticles generally have unique plasmon resonance effects. Metal nanoparticles such as gold, silver, and magnetism are widely used in the field of visual detection. A phage can be assembled with metal nanoparticles to form an organic–inorganic hybrid probe due to its nanometer-scale size and excellent modifiability. Due to the unique plasmon resonance effect of this composite probe, this technology can be used to visually detect objects of interest under a dark-field microscope. In summary, this review summarizes the recent advances in the development of phage-based probes for ultra-sensitive detection of various bio-species, outlining the advantages and limitations of detection technology of phage-based assays, and highlighting the commonly used editing technologies of phage genomes such as homologous recombination and clustered regularly interspaced palindromic repeats/CRISPR-associated proteins system (CRISPR-Cas). Finally, we discuss the possible scenarios for clinical application of phage-probe-based detection methods.

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“…Furthermore, the association between the core particle and large surface protein (L-HBsAg) during viral morphogenesis [17] was shown to be greatly inhibited by M13 phages that carry disulfide constrained heptapeptides, bearing the sequence C-WSFFSNI-C on their gpIII proteins [18]. These phages bearing the sequence C-WSFFSNI-C, have also exhibited great potential in detection of HBcAg in human serum as well as in precipitation of HBcAg from bacterial lysate [19]. Therefore, the aim of this study was to develop a method to purify HBcAg from unclarified Escherichia coli feedstock using M13 phage bearing the heptapeptides sequence, C-WSFFSNI-C, immobilized onto epoxidated Streamline base matrix via EBAC.…”

Section: Introductionmentioning

confidence: 97%