Efficient In Vivo Selection of a Novel Tumor-Associated Peptide from a Phage Display Library

Veleva, Anka N.; Nepal, Desh Bandhu; Frederick, C. Brandon; Schwab, Jacob; Lockyer, Pamela; Yuan, Hong; Lalush, David S.; Patterson, Cam

doi:10.3390/molecules16010900

Cited by 16 publications

(4 citation statements)

References 30 publications

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“…Peptides and peptide mimetics with high affinity and selectivity for biological targets will play a critical role in next generation medicinal chemistry, diagnostics, and downstream biomanufacturing technology 2,3,31–34 . Streamlining their identification requires affordable, automated devices capable of sorting solid-phase peptide library beads with accurate and rigorous orthogonal selection logics.…”

Section: Resultsmentioning

confidence: 99%

Affordable Microfluidic Bead-Sorting Platform for Automated Selection of Porous Particles Functionalized with Bioactive Compounds

Saberi-Bosari

Omary

Lavoie

et al. 2019

Sci Rep

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The ability to rapidly and accurately evaluate bioactive compounds immobilized on porous particles is crucial in the discovery of drugs, diagnostic reagents, ligands, and catalysts. Existing options for solid phase screening of bioactive compounds, while highly effective and well established, can be cost-prohibitive for proof-of-concept and early stage work, limiting its applicability and flexibility in new research areas. Here, we present a low-cost microfluidics-based platform enabling automated screening of small porous beads from solid-phase peptide libraries with high sensitivity and specificity, to identify leads with high binding affinity for a biological target. The integration of unbiased computer assisted image processing and analysis tools, provided the platform with the flexibility of sorting through beads with distinct fluorescence patterns. The customized design of the microfluidic device helped with handling beads with different diameters (~100–300 µm). As a microfluidic device, this portable novel platform can be integrated with a variety of analytical instruments to perform screening. In this study, the system utilizes fluorescence microscopy and unsupervised image analysis, and can operate at a sorting speed of up to 125 beads/hr (~3.5 times faster than a trained operator) providing >90% yield and >90% bead sorting accuracy. Notably, the device has proven successful in screening a model solid-phase peptide library by showing the ability to select beads carrying peptides binding a target protein (human IgG).

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Section: Resultsmentioning

confidence: 99%

Affordable Microfluidic Bead-Sorting Platform for Automated Selection of Porous Particles Functionalized with Bioactive Compounds

Saberi-Bosari

Omary

Lavoie

et al. 2019

Sci Rep

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“…Soendergaard et al used in vivo phage display selection to identify an ovarian cancer targeting peptide 46 . Another study of Veleva et al, applied a combination of in vitro and in vivo screening to isolate a peptide that is selective for circulating bone marrow derived cells from angiogenic Lung Lewis carcinoma tumors 47 . To the best of our knowledge, this is the first time that an in vivo selection has been applied in the selection of sdAbs against lymphoma and for ADC development.…”

Section: Discussionmentioning

confidence: 99%

Rabbit derived VL single-domains as promising scaffolds to generate antibody–drug conjugates

André

Dias

Aguiar

et al. 2023

Sci Rep

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Antibody–drug conjugates (ADCs) are among the fastest-growing classes of therapeutics in oncology. Although ADCs are in the spotlight, they still present significant engineering challenges. Therefore, there is an urgent need to develop more stable and effective ADCs. Most rabbit light chains have an extra disulfide bridge, that links the variable and constant domains, between Cys80 and Cys171, which is not found in the human or mouse. Thus, to develop a new generation of ADCs, we explored the potential of rabbit-derived VL-single-domain antibody scaffolds (sdAbs) to selectively conjugate a payload to Cys80. Hence, a rabbit sdAb library directed towards canine non-Hodgkin lymphoma (cNHL) was subjected to in vitro and in vivo phage display. This allowed the identification of several highly specific VL-sdAbs, including C5, which specifically target cNHL cells in vitro and present promising in vivo tumor uptake. C5 was selected for SN-38 site-selective payload conjugation through its exposed free Cys80 to generate a stable and homogenous C5-DAB-SN-38. C5-DAB-SN-38 exhibited potent cytotoxicity activity against cNHL cells while inhibiting DNA-TopoI activity. Overall, our strategy validates a platform to develop a novel class of ADCs that combines the benefits of rabbit VL-sdAb scaffolds and the canine lymphoma model as a powerful framework for clinically translation of novel therapeutics for cancer.

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“…This candidate was therefore selected to detect protease activity in tumors, directing therapeutic or imaging agents ( Whitney et al, 2010 ). Finally, Veleva et al (2011) group designed a strategy to identify ligands from a phage display random peptide library, selective for cells homing angiogenic tumors derived from circulating bone marrow, that included an in vitro phage display selection on blood outgrowth endothelial cells followed by an in vivo selection based on homing of bone marrow-bound phage to angiogenic tumors. This dual-functional strategy enabled the isolation of the peptide containing the QFPPKLTNNSML sequence.…”

Section: In Vivo Phage Displaymentioning

confidence: 99%

In vivo Phage Display: A promising selection strategy for the improvement of antibody targeting and drug delivery properties

et al. 2022

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The discovery of hybridoma technology, described by Kohler and Milstein in 1975, and the resulting ability to generate monoclonal antibodies (mAbs) initiated a new era in antibody research and clinical development. However, limitations of the hybridoma technology as a routine antibody generation method in conjunction with high immunogenicity responses have led to the development of alternative approaches for the streamlined identification of most effective antibodies. Within this context, display selection technologies such as phage display, ribosome display, yeast display, bacterial display, and mammalian cell surface display have been widely promoted over the past three decades as ideal alternatives to traditional hybridoma methods. The display of antibodies on phages is probably the most widespread and powerful of these methods and, since its invention in late 1980s, significant technological advancements in the design, construction, and selection of antibody libraries have been made, and several fully human antibodies generated by phage display are currently approved or in various clinical development stages. With evolving novel disease targets and the emerging of a new generation of therapeutic antibodies, such as bispecific antibodies, antibody drug conjugates (ADCs), and chimeric antigen receptor T (CAR-T) cell therapies, it is clear that phage display is expected to continue to play a central role in antibody development. Nevertheless, for non-standard and more demanding cases aiming to generate best-in-class therapeutic antibodies against challenging targets and unmet medical needs, in vivo phage display selections by which phage libraries are directly injected into animals or humans for isolating and identifying the phages bound to specific tissues offer an advantage over conventional in vitro phage display screening procedures. Thus, in the present review, we will first summarize a general overview of the antibody therapeutic market, the different types of antibody fragments, and novel engineered variants that have already been explored. Then, we will discuss the state-of-the-art of in vivo phage display methodologies as a promising emerging selection strategy for improvement antibody targeting and drug delivery properties.

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Efficient In Vivo Selection of a Novel Tumor-Associated Peptide from a Phage Display Library

Cited by 16 publications

References 30 publications

Affordable Microfluidic Bead-Sorting Platform for Automated Selection of Porous Particles Functionalized with Bioactive Compounds

Affordable Microfluidic Bead-Sorting Platform for Automated Selection of Porous Particles Functionalized with Bioactive Compounds

Rabbit derived VL single-domains as promising scaffolds to generate antibody–drug conjugates

In vivo Phage Display: A promising selection strategy for the improvement of antibody targeting and drug delivery properties

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