An in vivo selection-derived
            <scp>d</scp>
            -peptide for engineering erythrocyte-binding antigens that promote immune tolerance

Loftis, Alexander R.; Zhang, Genwei; Backlund, Coralie M.; Quartararo, Anthony J.; Pishesha, Novalia; Hanna, Cameron C.; Schissel, Carly K.; Garafola, Daniel; Loas, Andrei; Collier, R. J.; Ploegh, Hidde L.; Irvine, Darrell J.; Pentelute, Bradley L.

doi:10.1073/pnas.2101596118

Cited by 6 publications

(15 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Affinity selection–mass spectrometry (AS–MS) techniques have traditionally been used to probe protein–protein interactions in vitro . Our group has recently shown that chemical libraries may reach the diversity of other display techniques for identification of peptide binders to proteins, and that this strategy can be applied for cell-surface selection in vivo . In this work, we have added an additional spatial element to this strategy by extracting the cytosol for in-cell selection of fully synthetic peptide libraries conjugated to a model antisense cargo.…”

Section: Discussionmentioning

confidence: 99%

“…34 Our group has recently shown that chemical libraries may reach the diversity of other display techniques for identification of peptide binders to proteins, 33 and that this strategy can be applied for cell-surface selection in vivo. 42 In this work, we have added an additional spatial element to this strategy by extracting the cytosol for in-cell selection of fully synthetic peptide libraries conjugated to a model antisense cargo. By comparing these sequences to those found in a whole cell extract, we can exclude sequences that accumulate in the endosomes.…”

Section: ■ Conclusionmentioning

confidence: 99%

“…Our group has recently demonstrated that in vivo affinity selection−mass spectrometry (AS−MS) could identify an erythrocyte-targeting D-peptide. 42 Such labelfree techniques applied to the cell surface can be used to discover novel, non-canonical D-peptide binders without the addition of display scaffolds or encoding tags.…”

Section: ■ Introductionmentioning

confidence: 99%

See 2 more Smart Citations

In-Cell Penetration Selection–Mass Spectrometry Produces Noncanonical Peptides for Antisense Delivery

et al. 2023

Self Cite

View full text Add to dashboard Cite

Peptide-mediated delivery of macromolecules in cells has significant potential therapeutic benefits, but no therapy employing cell-penetrating peptides (CPPs) has reached the market after 30 years of investigation due to challenges in the discovery of new, more efficient sequences. Here, we demonstrate a method for in-cell penetration selection−mass spectrometry (in-cell PS−MS) to discover peptides from a synthetic library capable of delivering macromolecule cargo to the cytosol. This method was inspired by recent in vivo selection approaches for cellsurface screening, with an added spatial dimension resulting from subcellular fractionation. A representative peptide discovered in the cytosolic extract, Cyto1a, is nearly 100-fold more active toward antisense phosphorodiamidate morpholino oligomer (PMO) delivery compared to a sequence identified from a whole cell extract, which includes endosomes. Cyto1a is composed of Dresidues and two non-α-amino acids, is more stable than its all-L isoform, and is less toxic than known CPPs with comparable activity. Pulse-chase and microscopy experiments revealed that while the PMO−Cyto1a conjugate is likely taken up by endosomes, it can escape to localize to the nucleus without nonspecifically releasing other endosomal components. In-cell PS−MS introduces a means to empirically discover unnatural synthetic peptides for subcellular delivery of therapeutically relevant cargo.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: ■ Conclusionmentioning

confidence: 99%

See 1 more Smart Citation

In-Cell Penetration Selection–Mass Spectrometry Produces Noncanonical Peptides for Antisense Delivery

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

“…28 Our group has recently shown that chemical libraries may reach the diversity of other display techniques for identification of peptide binders to proteins 27 , and that this strategy can be applied for cell-surface selection in vivo. 36 In this work, we have added an additional spatial element to this strategy, by extracting the cytosol for in-cell selection of fully synthetic peptide libraries conjugated to a model antisense cargo. By comparing these sequences to those found in a whole cell extract, we can exclude sequences that accumulate in the endosomes.…”

Section: Discussionmentioning

confidence: 99%

“…Our group has recently demonstrated that in vivo affinity selection-mass spectrometry (AS-MS) could identify an erythrocyte-targeting D-peptide. 36 Such label-free techniques applied to the cell surface can be used to discover novel, non-canonical, D-peptide binders without the addition of display scaffolds or encoding tags.…”

Section: Introductionmentioning

confidence: 99%

In-cell penetration selection—mass spectrometry produces noncanonical peptides for antisense delivery

Schissel

Farquhar

Loas

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Peptide-mediated delivery of macromolecules in cells has significant potential therapeutic benefits, but no therapy employing cell-penetrating peptides (CPPs) has reached the market after 30 years of investigation due to challenges in the discovery of new, more efficient sequences. We developed a method for in-cell penetration selection-mass spectrometry (in-cell PS-MS) to discover peptides from a synthetic library capable of delivering macromolecule cargo to the cytosol. This method was inspired by recent in vivo selection approaches for cell-surface screening, with an added spatial dimension resulting from subcellular fractionation. A representative peptide discovered in the cytosolic extract, Pep1a, is nearly 100-fold more active toward antisense phosphorodiamidate morpholino oligomer (PMO) delivery compared to a sequence identified from a whole cell extract, which includes endosomes. Pep1a is composed of D-amino acids and two non-α-amino acids. Pulse-chase and microscopy experiments revealed that while the PMO-Pep1a conjugate is likely taken up by endosomes, it can escape to localize to the nucleus. In-cell PS-MS introduces a means to empirically discover unnatural synthetic peptides for subcellular delivery of therapeutically relevant cargo.

show abstract

Chiral Nanomaterials for Cancer Vaccines

Qu,

Sun,

et al. 2023

Small Methods

View full text Add to dashboard Cite

Chirality is a fundamental characteristic of living organisms and is commonly observed at the biomolecule, cellular, and tissue levels. Chiral nanomaterials play an irreplaceable role in nanomedicine and nanobiology because of their unique enantioselectivity with biological components. Here, research progress relating to chiral nanomaterials in the field of vaccines is reviewed, including antigen presenting systems, immune adjuvants, and cancer vaccines. First, the common synthesis methods are outlined for different types of chiral nanomaterials, as well as their chiral sources, optical properties, and potential biological applications. Then, the application of chiral nanomaterials are discussed in the field of vaccines with reference to the promotion of antigen presentation and activation of the immune system for tumor immunotherapy. Finally, the current obstacles and future research directions of chiral nanomaterials are revealed with regard to regulating the immune system.

show abstract

An in vivo selection-derived d -peptide for engineering erythrocyte-binding antigens that promote immune tolerance

Cited by 6 publications

References 36 publications

In-Cell Penetration Selection–Mass Spectrometry Produces Noncanonical Peptides for Antisense Delivery

In-Cell Penetration Selection–Mass Spectrometry Produces Noncanonical Peptides for Antisense Delivery

In-cell penetration selection—mass spectrometry produces noncanonical peptides for antisense delivery

Chiral Nanomaterials for Cancer Vaccines

Contact Info

Product

Resources

About