Identification and analysis of anti-HDL scFv-antibodies obtained from phage display based synthetic antibody library

Negi, Priyanka; Lövgren, Janita; Malmi, Päivi; Sirkka, Nina; Metso, Jari; Huovinen, Tuomas; Brockmann, Eeva-Christine; Pettersson, Kim; Jauhiainen, Matti; Lamminmäki, Urpo

doi:10.1016/j.clinbiochem.2015.11.020

Cited by 10 publications

(5 citation statements)

References 35 publications

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“…By using this technique, VHH single-domain antibodies produced from phage libraries can be recovered that recognize antigenic epitopes which could not be recognized by other previously tested agents. The construction of the phage antibody library depends on the high variability region of the conventional antibody and VHH recombinant antibody CDRs, which will determine whether the phage antibody will have a great relationship with the antibody library[ 38 ]. Conventional IgG antibodies have six antigen complementary binding regions (CDR), while VHH has only three antigen CDRs[ 39 , 40 ].…”

Section: Discussionmentioning

confidence: 99%

Selection and characterization of specific nanobody against bovine virus diarrhea virus (BVDV) E2 protein

Huang

Xiao

et al. 2017

PLoS ONE

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Bovine viral diarrhea-mucosal disease (BVD-MD) is caused by bovine viral diarrhea virus (BVDV), and results in abortion, stillbirth, and fetal malformation in cows. Here, we constructed the phage display vector pCANTAB 5E-VHH and then transformed it into Escherichia coli TG1-competent cells, to construct an initial anti-BVDV nanobody gene library. We obtained a BVDV-E2 antigen epitope bait protein by prokaryotic expression using the nucleotide sequence of the E2 gene of the BVDV-NADL strain published in GenBank. Phage display was used to screen the anti-BVDV nanobody gene library. We successfully constructed a high quality phage display nanobody library, with an initial library capacity of 4.32×105. After the rescue of helper phage, the titer of the phage display nanobody library was 1.3×1011. The BVDV-E2 protein was then expressed in Escherichia coli (DE3), and a 49.5 kDa band was observed with SDS-PAGE analysis that was consistent with the expected nanobody size. Thus, we were able to isolate one nanobody that exhibits high affinity and specificity against BVDV using phage display techniques. This isolated nanobody was then used in Enzyme Linked Immunosorbent Assay and qRT-PCR, and ELISA analyses of BVDV infection of MDBK cells indicated that the nanobodies exhibited good antiviral effect.

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Section: Discussionmentioning

confidence: 99%

Selection and characterization of specific nanobody against bovine virus diarrhea virus (BVDV) E2 protein

Huang

Xiao

et al. 2017

PLoS ONE

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“…After 20 h induction at 30 °C, cells were harvested by centrifugation, resuspended in PBS and disrupted by sonication. The scFv protein was purified by immobilized metal affinity chromatography (IMAC), followed by a Superdex 75 size-exclusion chromatography using PBS as running buffer in a Biologic (Bio-Rad) as described previously [ 35 ]. Protein concentrations were quantified with the micro-BCA kit (Pierce, Thermo Scientific).…”

Section: Methodsmentioning

confidence: 99%

A Novel Fully Human Agonistic Single Chain Fragment Variable Antibody Targeting Death Receptor 5 with Potent Antitumor Activity In Vitro and In Vivo

Lei¹,

Xu²,

Xu³

et al. 2017

IJMS

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Agonistic antibodies, which bind specifically to death receptor 5 (DR5), can trigger apoptosis in tumor cells through the extrinsic pathway. In this present study, we describe the use of a phage display to isolate a novel fully human agonistic single chain fragment variable (scFv) antibody, which targets DR5. After five rounds of panning a large (1.2 × 108 clones) phage display library on DR5, a total of over 4000 scFv clones were screened by the phage ELISA. After screening for agonism in a cell-viability assay in vitro, a novel DR5-specific scFv antibody TR2-3 was isolated, which inhibited COLO205 and MDA-MB-231 tumor cell growth without any cross-linking agents. The activity of TR2-3 in inducing apoptosis in cancer cells was evaluated by using an Annexin V-PE apoptosis detection kit in combination with flow cytometry and the Hoechst 33342 and propidium iodide double staining analysis. In addition, the activation of caspase-dependent apoptosis was evaluated by Western blot assays. The results indicated that TR2-3 induced robust apoptosis of the COLO205 and MDA-MB-231 cells in a dose-dependent and time-dependent manner, while it remarkably upregulated the cleavage of caspase-3 and caspase-8. Furthermore, TR2-3 suppressed the tumor growth significantly in the xenograft model. Taken together, these data suggest that TR2-3 exhibited potent antitumor activity both in vitro and in vivo. This work provides a novel human antibody, which might be a promising candidate for cancer therapy by targeting DR5.

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“…Moreover, anti-HDL (high-density lipoprotein) antibodies have been produced through the biopanning technique using HDL derived from patients diagnosed with coronary artery disease. The ability of these antibodies to bind to apolipoproteins A-1 and 2 demonstrates the diagnostic utility of the phage display technology [102].…”

Section: Cardiovascular Diseasesmentioning

confidence: 97%

Phage Display Technology in Biomarker Identification with Emphasis on Non-Cancerous Diseases

Sadraeian,

Maleki,

Moraghebi

et al. 2024

Molecules

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In recent years, phage display technology has become vital in clinical research. It helps create antibodies that can specifically bind to complex antigens, which is crucial for identifying biomarkers and improving diagnostics and treatments. However, existing reviews often overlook its importance in areas outside cancer research. This review aims to fill that gap by explaining the basics of phage display and its applications in detecting and treating various non-cancerous diseases. We focus especially on its role in degenerative diseases, inflammatory and autoimmune diseases, and chronic non-communicable diseases, showing how it is changing the way we diagnose and treat illnesses. By highlighting important discoveries and future possibilities, we hope to emphasize the significance of phage display in modern healthcare.

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Identification and analysis of anti-HDL scFv-antibodies obtained from phage display based synthetic antibody library

Cited by 10 publications

References 35 publications

Selection and characterization of specific nanobody against bovine virus diarrhea virus (BVDV) E2 protein

Selection and characterization of specific nanobody against bovine virus diarrhea virus (BVDV) E2 protein

A Novel Fully Human Agonistic Single Chain Fragment Variable Antibody Targeting Death Receptor 5 with Potent Antitumor Activity In Vitro and In Vivo

Phage Display Technology in Biomarker Identification with Emphasis on Non-Cancerous Diseases

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