Construction of Synthetic Antibody Phage Display Libraries

Giang, Kim Anh; Sidhu, Sachdev S.; Nilvebrant, Johan

doi:10.1007/978-1-0716-3381-6_4

Cited by 2 publications

(3 citation statements)

References 43 publications

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“…Phage precipitation from LB medium with PEG/NaCl (5:1 ratio between LB medium and PEG/NaCl) on ice for 1 h was followed by centrifugation at 4600 rpm for 15 min at 4 °C to pellet phages. Phages were resuspended in sterile PBS and phage titer was determined by measuring absorption at 269 and 320 nm on Nanodrop (Thermo Fisher Scientific, Waltham, MA, USA) and calculated as followed: virions/ml = (A269-A320)*6*10 16 /number of bases per virion 35 , 36 .…”

Section: Methodsmentioning

confidence: 99%

In vivo phage display identifies novel peptides for cardiac targeting

Ivanova,

Kohl,

González-King Garibotti

et al. 2024

Sci Rep

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Heart failure remains a leading cause of mortality. Therapeutic intervention for heart failure would benefit from targeted delivery to the damaged heart tissue. Here, we applied in vivo peptide phage display coupled with high-throughput Next-Generation Sequencing (NGS) and identified peptides specifically targeting damaged cardiac tissue. We established a bioinformatics pipeline for the identification of cardiac targeting peptides. Hit peptides demonstrated preferential uptake by human induced pluripotent stem cell (iPSC)-derived cardiomyocytes and immortalized mouse HL1 cardiomyocytes, without substantial uptake in human liver HepG2 cells. These novel peptides hold promise for use in targeted drug delivery and regenerative strategies and open new avenues in cardiovascular research and clinical practice.

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Section: Methodsmentioning

confidence: 99%

In vivo phage display identifies novel peptides for cardiac targeting

Ivanova,

Kohl,

González-King Garibotti

et al. 2024

Sci Rep

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show abstract

“…A plausible alternative that allows for faster production of mAbs is through in vitro-directed protein evolution procedures, which involve (1) the generation of native and immune gene libraries [ 12 ] or synthetic [ 13 ] that encode single-chain variable fragment ( scFv ) mAbs displayed through phage (phage display) [ 14 ]; (2) the process of screening and selection of isoforms related to an immobilized antigen (target proteins and/or previously identified biomarkers) [ 15 ]; and (3) the in vitro maturation of the affinity or specificity of the antibody, through the insertion of random mutations and selection of the isoform with improved affinity [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

“…These antibodies are fused with proteins from the phage coat and displayed on its surface, facilitating the isolation of scFv and Fab fragments targeting specific cellular markers [ 14 ]. Antibody libraries can be generated from pre-immune donors using V domains from the germ line or synthetic genes [ 12 , 13 ]. The multivalent presentation of antibodies is crucial for efficient selection.…”

Section: Introductionmentioning

confidence: 99%

Construction of a Human Immune Library from Gallbladder Cancer Patients for the Single-Chain Fragment Variable (scFv) Antibody Selection against Claudin 18.2 via Phage Display

Effer,

Ulloa,

Dappolonnio

et al. 2024

Antibodies

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Gallbladder cancer (GBC) is a very aggressive malignant neoplasm of the biliary tract with a poor prognosis. There are no specific therapies for the treatment of GBC or early diagnosis tools; for this reason, the development of strategies and technologies that facilitate or allow an early diagnosis of GBC continues to be decisive. Phage display is a robust technique used for the production of monoclonal antibodies (mAbs) involving (1) the generation of gene libraries, (2) the screening and selection of isoforms related to an immobilized antigen, and (3) the in vitro maturation of the affinity of the antibody for the antigen. This research aimed to construct a human immune library from PBMCs of GBC patients and the isolation of scFv-phage clones with specificity against the larger extracellular loop belonging to claudin 18.2, which is an important biomarker overexpressed in GBC as well as gastric cancer. The immune-library-denominated GALLBLA1 was constructed from seven GBC patients and has a diversity of 6.12 × 1010 pfu mL−1. After three rounds of panning, we were able to identify clones with specificity against claudin 18.2. GALLBLA1 can contribute to the selection, isolation, and recombinant production of new human mAbs candidates for the treatment of gastrointestinal cancers.

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Construction of Synthetic Antibody Phage Display Libraries

Cited by 2 publications

References 43 publications

In vivo phage display identifies novel peptides for cardiac targeting

In vivo phage display identifies novel peptides for cardiac targeting

Construction of a Human Immune Library from Gallbladder Cancer Patients for the Single-Chain Fragment Variable (scFv) Antibody Selection against Claudin 18.2 via Phage Display

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