Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation. / Kristensen, Mie; de Groot, Anne Marit; Berthelsen, Jens; Franzyk, Henrik; Sijts, Alice; Nielsen, Hanne Mørck.

In: Bioconjugate Chemistry, Vol. 26, 2015, p. 477-488.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Kristensen, M, de Groot, AM, Berthelsen, J, Franzyk, H, Sijts, A & Nielsen, HM 2015, 'Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation', Bioconjugate Chemistry, vol. 26, pp. 477-488. https://doi.org/10.1021/bc5005763

APA

Kristensen, M., de Groot, A. M., Berthelsen, J., Franzyk, H., Sijts, A., & Nielsen, H. M. (2015). Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation. Bioconjugate Chemistry, 26, 477-488. https://doi.org/10.1021/bc5005763

Vancouver

Kristensen M, de Groot AM, Berthelsen J, Franzyk H, Sijts A, Nielsen HM. Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation. Bioconjugate Chemistry. 2015;26:477-488. https://doi.org/10.1021/bc5005763

Author

Kristensen, Mie ; de Groot, Anne Marit ; Berthelsen, Jens ; Franzyk, Henrik ; Sijts, Alice ; Nielsen, Hanne Mørck. / Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation. In: Bioconjugate Chemistry. 2015 ; Vol. 26. pp. 477-488.

Bibtex

@article{d2127f6e04c64bf6852e0218647bc1dc,
title = "Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation",
abstract = "Delivery of therapeutic peptides and proteins by the use of cell-penetrating peptides (CPPs) as carriers has been suggested as a feasible strategy. The aim of the present study was to investigate the effect of conjugating a series of well-known CPPs to the biologically active part of parathyroid hormone, i.e. PTH(1-34), and to evaluate the effect with regards to secondary structure, potency in Saos-2 cells, immunogenicity, safety as well as the transepithelial permeation across monolayers by using the Caco-2 cell culture model. Further, co-administration of CPP and PTH(1-34) as an alternative to covalent conjugation was compared with regards to the transepithelial permeation. CPP-conjugated PTH(1-34) fusion peptides were successfully expressed in Escherichia coli and purified from inclusion bodies. No clear correlation between the degree of secondary structure of the CPP-conjugated PTH(1-34) fusion peptides and their potency was found, albeit a general decrease in permeation was observed for both N- and C-terminally CPP-conjugated PTH(1-34) as compared to native PTH(1-34). However, attachment of CPP to the N-terminus significantly increased permeation across Caco-2 cell monolayers as compared to the corresponding C-terminally CPP-conjugated PTH(1-34). In addition, the nonaarginine sequence proved to be the only CPP capable of increasing permeation when conjugated to PTH(1-34) as compared to co-administration of CPP and PTH(1-34). This enhancement effect was, however, associated with an unacceptable low level of cell viability. In conclusion, covalent conjugation of CPPs to PTH(1-34) influenced both the secondary structure, potency, and transepithelial permeation efficiency of the resulting conjugate, and hence this approach appears not to be favorable as compared to co-administration when optimizing CPP-mediated permeation of PTH(1-34) across an intestinal epithelium.",
author = "Mie Kristensen and {de Groot}, {Anne Marit} and Jens Berthelsen and Henrik Franzyk and Alice Sijts and Nielsen, {Hanne M{\o}rck}",
year = "2015",
doi = "10.1021/bc5005763",
language = "English",
volume = "26",
pages = "477--488",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",

}

RIS

TY - JOUR

T1 - Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation

AU - Kristensen, Mie

AU - de Groot, Anne Marit

AU - Berthelsen, Jens

AU - Franzyk, Henrik

AU - Sijts, Alice

AU - Nielsen, Hanne Mørck

PY - 2015

Y1 - 2015

N2 - Delivery of therapeutic peptides and proteins by the use of cell-penetrating peptides (CPPs) as carriers has been suggested as a feasible strategy. The aim of the present study was to investigate the effect of conjugating a series of well-known CPPs to the biologically active part of parathyroid hormone, i.e. PTH(1-34), and to evaluate the effect with regards to secondary structure, potency in Saos-2 cells, immunogenicity, safety as well as the transepithelial permeation across monolayers by using the Caco-2 cell culture model. Further, co-administration of CPP and PTH(1-34) as an alternative to covalent conjugation was compared with regards to the transepithelial permeation. CPP-conjugated PTH(1-34) fusion peptides were successfully expressed in Escherichia coli and purified from inclusion bodies. No clear correlation between the degree of secondary structure of the CPP-conjugated PTH(1-34) fusion peptides and their potency was found, albeit a general decrease in permeation was observed for both N- and C-terminally CPP-conjugated PTH(1-34) as compared to native PTH(1-34). However, attachment of CPP to the N-terminus significantly increased permeation across Caco-2 cell monolayers as compared to the corresponding C-terminally CPP-conjugated PTH(1-34). In addition, the nonaarginine sequence proved to be the only CPP capable of increasing permeation when conjugated to PTH(1-34) as compared to co-administration of CPP and PTH(1-34). This enhancement effect was, however, associated with an unacceptable low level of cell viability. In conclusion, covalent conjugation of CPPs to PTH(1-34) influenced both the secondary structure, potency, and transepithelial permeation efficiency of the resulting conjugate, and hence this approach appears not to be favorable as compared to co-administration when optimizing CPP-mediated permeation of PTH(1-34) across an intestinal epithelium.

AB - Delivery of therapeutic peptides and proteins by the use of cell-penetrating peptides (CPPs) as carriers has been suggested as a feasible strategy. The aim of the present study was to investigate the effect of conjugating a series of well-known CPPs to the biologically active part of parathyroid hormone, i.e. PTH(1-34), and to evaluate the effect with regards to secondary structure, potency in Saos-2 cells, immunogenicity, safety as well as the transepithelial permeation across monolayers by using the Caco-2 cell culture model. Further, co-administration of CPP and PTH(1-34) as an alternative to covalent conjugation was compared with regards to the transepithelial permeation. CPP-conjugated PTH(1-34) fusion peptides were successfully expressed in Escherichia coli and purified from inclusion bodies. No clear correlation between the degree of secondary structure of the CPP-conjugated PTH(1-34) fusion peptides and their potency was found, albeit a general decrease in permeation was observed for both N- and C-terminally CPP-conjugated PTH(1-34) as compared to native PTH(1-34). However, attachment of CPP to the N-terminus significantly increased permeation across Caco-2 cell monolayers as compared to the corresponding C-terminally CPP-conjugated PTH(1-34). In addition, the nonaarginine sequence proved to be the only CPP capable of increasing permeation when conjugated to PTH(1-34) as compared to co-administration of CPP and PTH(1-34). This enhancement effect was, however, associated with an unacceptable low level of cell viability. In conclusion, covalent conjugation of CPPs to PTH(1-34) influenced both the secondary structure, potency, and transepithelial permeation efficiency of the resulting conjugate, and hence this approach appears not to be favorable as compared to co-administration when optimizing CPP-mediated permeation of PTH(1-34) across an intestinal epithelium.

U2 - 10.1021/bc5005763

DO - 10.1021/bc5005763

M3 - Journal article

VL - 26

SP - 477

EP - 488

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

ER -

ID: 130643656