Detection of a prostate cancer cell line using a bioluminescent affiprobe: An attempt to develop a new molecular probe for ex vivo studies

Varnosfaderani, Zahra Ghafouri; Emamzadeh, Rahman; Nazari, Mahboobeh; Zarean, Mojtaba

doi:10.1016/j.ijbiomac.2019.07.085

Cited by 8 publications

(4 citation statements)

References 49 publications

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“…As a sensory component of hybrids, various biospecific polypeptides were applied—antibodies or their fragments [ 111 , 112 , 113 , 114 ], antigens [ 115 , 116 , 117 , 118 ], polypeptides with special binding ability [ 119 , 120 , 121 , 122 ], GFP and its color variants [ 123 , 124 , 125 , 126 , 127 , 128 , 129 ], etc. Based on the phenomenon of assembling split luciferase derivatives, a whole spectrum of homogeneous methods has been developed [ 125 , 126 , 127 , 128 , 129 ] and applied for studying protein–protein/ligand interaction [ 130 , 131 , 132 , 133 ], protein activity [ 134 , 135 ], and protein folding [ 136 , 137 ].…”

Section: Ctz-dependent Luciferase Analytical Applicationmentioning

confidence: 99%

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Krasitskaya

Bashmakova

Frank

2020

IJMS

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: The functioning of bioluminescent systems in most of the known marine organisms is based on the oxidation reaction of the same substrate—coelenterazine (CTZ), catalyzed by luciferase. Despite the diversity in structures and the functioning mechanisms, these enzymes can be united into a common group called CTZ-dependent luciferases. Among these, there are two sharply different types of the system organization—Ca2+-regulated photoproteins and luciferases themselves that function in accordance with the classical enzyme–substrate kinetics. Along with deep and comprehensive fundamental research on these systems, approaches and methods of their practical use as highly sensitive reporters in analytics have been developed. The research aiming at the creation of artificial luciferases and synthetic CTZ analogues with new unique properties has led to the development of new experimental analytical methods based on them. The commercial availability of many ready-to-use assay systems based on CTZ-dependent luciferases is also important when choosing them by first-time-users. The development of analytical methods based on these bioluminescent systems is currently booming. The bioluminescent systems under consideration were successfully applied in various biological research areas, which confirms them to be a powerful analytical tool. In this review, we consider the main directions, results, and achievements in research involving these luciferases.

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Section: Ctz-dependent Luciferase Analytical Applicationmentioning

confidence: 99%

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Krasitskaya

Bashmakova

Frank

2020

IJMS

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“…These signaling pathways regulate cell proliferation, survival, migration, angiogenesis, and metastasis. , However, Her2 is specifically overexpressed in many types of malignant tumors, including breast, gastric, and ovarian. − Her2 overexpression enhances cell signal transduction and accelerates the growth and metastasis of tumors, resulting in faster disease progression and poor clinical prognosis for patients bearing Her2-positive tumors . Therefore, Her2 is considered a diagnostic and treatment target for these tumors …”

Section: Introductionmentioning

confidence: 99%

“…11 Therefore, Her2 is considered a diagnostic and treatment target for these tumors. 12 Over the past decades, Her2-targeted therapy based on humanized monoclonal antibodies, including trastuzumab and pertuzumab, has achieved satisfactory outcomes and improved the survival rate of patients. 13,14 However, patients with latestage tumors always have poor treatment outcomes owing to untimely diagnosis and treatment.…”

Section: Introductionmentioning

confidence: 99%

ICG-Dimeric Her2-Specific Affibody Conjugates for Tumor Imaging and Photothermal Therapy for Her2-Positive Tumors

et al. 2022

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Human epidermal growth factor receptor 2 (Her2) is abundantly expressed in various solid tumors. The Her2specific Affibody (Z Her2:2891 ) has been clinically tested in patients with Her2-positive breast cancer and is regarded as an ideal drug carrier for tumor diagnosis and targeted treatment. Indocyanine green (ICG) can be used as a photosensitizer for photothermal therapy (PTT), in addition to fluorescent dyes for tumor imaging. In this study, a dimeric Her2-specific Affibody (Z Her2 ) based on Z Her2:2891 was prepared using the E. coli expression system and then coupled to ICG through an N-hydroxysuccinimide (NHS) ester reactive group to construct a novel bifunctional protein drug (named ICG-Z Her2 ) for tumor diagnosis and PTT. In vitro, ICG-Z Her2mediated PTT selectively and efficiently killed Her2-positive BT-474 and SKOV-3 tumor cells rather than Her2-negative HeLa tumor cells. In vivo, ICG-Z Her2 specifically accumulated in Her2-positive SKOV-3 tumor grafts rather than Her2-negative HeLa tumor grafts; high-contrast tumor optical images were obtained. However, Her2-negative HeLa tumor grafts were not detected. More importantly, ICG-Z Her2 -mediated PTT exhibited a significantly enhanced antitumor effect in mice bearing SKOV-3 tumor grafts owing to the good photothermal properties of ICG-Z Her2 . Of note, ICG-Z Her2 did not exhibit acute toxicity in mice during short-term treatment. Overall, our findings indicate that ICG-Z Her2 is a promising bifunctional drug for Her2-positive tumor diagnosis and PTT.

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“…2017; Varnosfaderani et al . 2019). Among them, GFP and enhanced green fluorescent protein (EGFP), a manipulated form of GFP, are stabile proteins in a variety of conditions such as in the presence of detergents, proteases and guanidinium chloride (GdmCl) (Stepanenko et al .…”

Section: Introductionmentioning

confidence: 99%

Novel luminescent affiprobes for molecular detection of Staphylococcus aureus using flow cytometry

2020

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Aims Diagnosis of Staphylococcus aureus is important in various diseases from hospital‐acquired infections to foodborne diseases. This work reports two new luminescent affiprobes for specific detection of S. aureus. Methods and Results To develop advanced luminescent affiprobes, enhanced green fluorescent protein (EGFP) was flanked by single and double repeats of ZpA963 affibody using molecular biology studies. The recombinant proteins including fluorescent monomeric affibody (fA1) and fluorescent dimeric affibody (fA2) were expressed in the bacterial expression system, purified and used to identify the S. aureus. Fluorescence microscope and flow cytometry results demonstrated that the treated samples with fA1 and fA2 had relatively high fluorescent mean intensities in comparison to the untreated S. aureus cells. Moreover, it was revealed that ‘fA2’ affiprobe had lower dissociation constant value (about 25‐fold) and was more effective for detection of S. aureus than the ‘fA1’ affiprobe. In addition, the binding of the affiprobes for some other pathogenic bacteria i.e. Escherichia coli, Bacillus cereus, Enterococcus faecalis and Staphylococcus saprophyticus was examined. Expectedly, no cross‐reaction was observed for binding the constructed affiprobes to these bacteria, eliminating possibilities for false positive results. Conclusions The results show that ‘fA1’ affiprobe and ‘fA2’ affiprobe are two new efficient luminescent affiprobes for detecting S. aureus. Significance and Impact of the Study We developed a new approach for detection of Staphylococcus aureus in a simple one‐step process and in low concentrations of probes. In the best of our knowledge, this is the first study to direct detection of bacterial cells by affiprobes and may be used to develop new diagnostic kits.

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Detection of a prostate cancer cell line using a bioluminescent affiprobe: An attempt to develop a new molecular probe for ex vivo studies

Cited by 8 publications

References 49 publications

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

ICG-Dimeric Her2-Specific Affibody Conjugates for Tumor Imaging and Photothermal Therapy for Her2-Positive Tumors

Novel luminescent affiprobes for molecular detection of Staphylococcus aureus using flow cytometry

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