Selection and characterization of cell binding and internalizing phage antibodies

Zhou, Yu; Zhao, Lifeng; Marks, James D.

doi:10.1016/j.abb.2012.05.007

Cited by 18 publications

(17 citation statements)

References 55 publications

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“…Internalizing antibodies can be obtained via direct selection of internalizing phage antibodies by incubating phage libraries directly with the target cells [575][576][577][578]. However, the major challenge in this process is to identify the antigen bound by the internalizing antibody, which can be determined indirectly using the flow cytometry of target cells, and identifying the cognate antigen recognized by tumor-specific antibodies using immunoprecipitation and mass spectrometry [579].…”

Section: Intracellular Targetingmentioning

confidence: 99%

Antibody Structure and Function: The Basis for Engineering Therapeutics

et al. 2019

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Antibodies and antibody-derived macromolecules have established themselves as the mainstay in protein-based therapeutic molecules (biologics). Our knowledge of the structure–function relationships of antibodies provides a platform for protein engineering that has been exploited to generate a wide range of biologics for a host of therapeutic indications. In this review, our basic understanding of the antibody structure is described along with how that knowledge has leveraged the engineering of antibody and antibody-related therapeutics having the appropriate antigen affinity, effector function, and biophysical properties. The platforms examined include the development of antibodies, antibody fragments, bispecific antibody, and antibody fusion products, whose efficacy and manufacturability can be improved via humanization, affinity modulation, and stability enhancement. We also review the design and selection of binding arms, and avidity modulation. Different strategies of preparing bispecific and multispecific molecules for an array of therapeutic applications are included.

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Section: Intracellular Targetingmentioning

confidence: 99%

Antibody Structure and Function: The Basis for Engineering Therapeutics

et al. 2019

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“…Oftentimes it is possible to enhance cellular internalization by antibody multimerization that can drive clustering of cell surface targets . Thus, internalization assays were performed using the RBE4 rat brain endothelial cell line and the internalization of biotinylated scFvA‐Avitag was compared to that of SEC‐purified scFvA‐tetramers.…”

Section: Resultsmentioning

confidence: 99%

Creation and evaluation of a single‐chain antibody tetramer that targets brain endothelial cells

2014

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Antibodies that target and internalize into blood-brain barrier (BBB) endothelial cells offer promise as drug delivery agents. Previously, we identified a single-chain antibody (scFvA) capable of binding to the BBB. In an attempt to improve the binding and internalization properties of the single chain antibody (scFvA), a biotinylation tag (Avitag) was fused to scFvA and the protein secreted by yeast. The scFvA-Avitag could be biotinylated by yeast-displayed BirA enzyme and biotinylated scFvA-Avitag could be used to create scFv tetramers. Tetramerization of scFvA improved the internalization of scFvA into BBB endothelial cells, and biotinylated scFvA-Avitag could also be used to target streptavidin-coated quantum dots for BBB endothelial cell internalization. Perfusing the rat brain with scFvA-tetramer confirmed that the antigen targeted by scFvA is distributed on blood side of the BBB, suggesting the potential for downstream application of scFvA in brain-targeted drug delivery.

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“…VCAM-1 binding peptides undergoing enhanced endocytosis provided improved imaging of vascular inflammation in animal models [190,191]. Phage display and other high-throughput methods facilitate selection of internalizable ligands [197–199]. …”

Section: Intracellular Endothelial Deliverymentioning

confidence: 99%

Targeted endothelial nanomedicine for common acute pathological conditions

Shuvaev

Brenner

Muzykantov

2015

Journal of Controlled Release

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Endothelium, a thin monolayer of specialized cells lining the lumen of blood vessels is the key regulatory interface between blood and tissues. Endothelial abnormalities are implicated in many diseases, including common acute conditions with high morbidity and mortality lacking therapy, in part because drugs and drug carriers have no natural endothelial affinity. Precise endothelial drug delivery may improve management of these conditions. Using ligands of molecules exposed to the bloodstream on the endothelial surface enables design of diverse targeted endothelial nanomedicine agents. Target molecules and binding epitopes must be accessible to drug carriers, carriers must be free of harmful effects, and targeting should provide desirable sub-cellular addressing of the drug cargo. The roster of current candidate target molecules for endothelial nanomedicine includes peptidases and other enzymes, cell adhesion molecules and integrins, localized in different domains of the endothelial plasmalemma and differentially distributed throughout the vasculature. Endowing carriers with an affinity to specific endothelial epitopes enables an unprecedented level of precision of control of drug delivery: binding to selected endothelial cell phenotypes, cellular addressing and duration of therapeutic effects. Features of nanocarrier design such as choice of epitope and ligand control delivery and effect of targeted endothelial nanomedicine agents. Pathological factors modulate endothelial targeting and uptake of nanocarriers. Selection of optimal binding sites and design features of nanocarriers are key controllable factors that can be iteratively engineered based on their performance from in vitro to pre-clinical in vivo experimental models. Targeted endothelial nanomedicine agents provide antioxidant, anti-inflammatory and other therapeutic effects unattainable by non-targeted counterparts in animal models of common acute severe human disease conditions. The results of animal studies provide the basis for the challenging translation endothelial nanomedicine into the clinical domain.

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Selection and characterization of cell binding and internalizing phage antibodies

Cited by 18 publications

References 55 publications

Antibody Structure and Function: The Basis for Engineering Therapeutics

Antibody Structure and Function: The Basis for Engineering Therapeutics

Creation and evaluation of a single‐chain antibody tetramer that targets brain endothelial cells

Targeted endothelial nanomedicine for common acute pathological conditions

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