Site-Specific 64Cu Labeling of the Serine Protease, Active Site Inhibited Factor Seven Azide (FVIIai-N3), Using Copper Free Click Chemistry

Troels E. Jeppesen; Lotte K. Kristensen; Carsten H. Nielsen; Lars C. Petersen; Jesper B. Kristensen; Carsten Behrens; Jacob Madsen; Andreas Kjaer

doi:10.1021/acs.bioconjchem.7b00649

Site-Specific ⁶⁴Cu Labeling of the Serine Protease, Active Site Inhibited Factor Seven Azide (FVIIai-N₃), Using Copper Free Click Chemistry

Troels E. Jeppesen, Lotte K. Kristensen, Carsten H. Nielsen, Lars C. Petersen, Jesper B. Kristensen, Carsten Behrens, Jacob Madsen, Andreas Kjaer^*

^*Corresponding author af dette arbejde

2 Citationer (Scopus)

Abstract

A method for site-specific radiolabeling of the serine protease active site inhibited factor seven (FVIIai) with ⁶⁴Cu has been applied using a biorthogonal click reaction. FVIIai binds to tissue factor (TF), a trans-membrane protein involved in hemostasis, angiogenesis, proliferation, cell migration, and survival of cancer cells. First a single azide moiety was introduced in the active site of this 50 kDa protease. Then a NOTA moiety was introduced via a strain promoted azide-alkyne reaction and the corresponding conjugate was labeled with ⁶⁴Cu. Binding to TF and the stability was evaluated in vitro. TF targeting capability of the radiolabeled conjugate was tested in vivo by positron emission tomography (PET) imaging in pancreatic human xenograft cancer mouse models with various TF expressions. The conjugate showed good stability (>91% at 16 h), an immunoreactivity of 93.5%, and a mean tumor uptake of 2.1 ± 0.2%ID/g at 15 h post injection. In conclusion, FVIIai was radiolabeled with ⁶⁴Cu in single well-defined position of the protein. This method can be utilized to prepare conjugates from serine proteases with the label at a specific position.

Originalsprog	Engelsk
Tidsskrift	Bioconjugate Chemistry
Vol/bind	29
Udgave nummer	1
Sider (fra-til)	117-125
Antal sider	9
ISSN	1043-1802
DOI	https://doi.org/10.1021/acs.bioconjchem.7b00649
Status	Udgivet - 2018

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10.1021/acs.bioconjchem.7b00649

Citationsformater

@article{5b916b21581c42bc8bb1278f8e46746e,

title = "Site-Specific 64Cu Labeling of the Serine Protease, Active Site Inhibited Factor Seven Azide (FVIIai-N3), Using Copper Free Click Chemistry",

abstract = "A method for site-specific radiolabeling of the serine protease active site inhibited factor seven (FVIIai) with 64Cu has been applied using a biorthogonal click reaction. FVIIai binds to tissue factor (TF), a trans-membrane protein involved in hemostasis, angiogenesis, proliferation, cell migration, and survival of cancer cells. First a single azide moiety was introduced in the active site of this 50 kDa protease. Then a NOTA moiety was introduced via a strain promoted azide-alkyne reaction and the corresponding conjugate was labeled with 64Cu. Binding to TF and the stability was evaluated in vitro. TF targeting capability of the radiolabeled conjugate was tested in vivo by positron emission tomography (PET) imaging in pancreatic human xenograft cancer mouse models with various TF expressions. The conjugate showed good stability (>91% at 16 h), an immunoreactivity of 93.5%, and a mean tumor uptake of 2.1 ± 0.2%ID/g at 15 h post injection. In conclusion, FVIIai was radiolabeled with 64Cu in single well-defined position of the protein. This method can be utilized to prepare conjugates from serine proteases with the label at a specific position.",

author = "Jeppesen, {Troels E.} and Kristensen, {Lotte K.} and Nielsen, {Carsten H.} and Petersen, {Lars C.} and Kristensen, {Jesper B.} and Carsten Behrens and Jacob Madsen and Andreas Kjaer",

year = "2018",

doi = "10.1021/acs.bioconjchem.7b00649",

language = "English",

volume = "29",

pages = "117--125",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

number = "1",

}

TY - JOUR

T1 - Site-Specific 64Cu Labeling of the Serine Protease, Active Site Inhibited Factor Seven Azide (FVIIai-N3), Using Copper Free Click Chemistry

AU - Jeppesen, Troels E.

AU - Kristensen, Lotte K.

AU - Nielsen, Carsten H.

AU - Petersen, Lars C.

AU - Kristensen, Jesper B.

AU - Behrens, Carsten

AU - Madsen, Jacob

AU - Kjaer, Andreas

PY - 2018

Y1 - 2018

N2 - A method for site-specific radiolabeling of the serine protease active site inhibited factor seven (FVIIai) with 64Cu has been applied using a biorthogonal click reaction. FVIIai binds to tissue factor (TF), a trans-membrane protein involved in hemostasis, angiogenesis, proliferation, cell migration, and survival of cancer cells. First a single azide moiety was introduced in the active site of this 50 kDa protease. Then a NOTA moiety was introduced via a strain promoted azide-alkyne reaction and the corresponding conjugate was labeled with 64Cu. Binding to TF and the stability was evaluated in vitro. TF targeting capability of the radiolabeled conjugate was tested in vivo by positron emission tomography (PET) imaging in pancreatic human xenograft cancer mouse models with various TF expressions. The conjugate showed good stability (>91% at 16 h), an immunoreactivity of 93.5%, and a mean tumor uptake of 2.1 ± 0.2%ID/g at 15 h post injection. In conclusion, FVIIai was radiolabeled with 64Cu in single well-defined position of the protein. This method can be utilized to prepare conjugates from serine proteases with the label at a specific position.

AB - A method for site-specific radiolabeling of the serine protease active site inhibited factor seven (FVIIai) with 64Cu has been applied using a biorthogonal click reaction. FVIIai binds to tissue factor (TF), a trans-membrane protein involved in hemostasis, angiogenesis, proliferation, cell migration, and survival of cancer cells. First a single azide moiety was introduced in the active site of this 50 kDa protease. Then a NOTA moiety was introduced via a strain promoted azide-alkyne reaction and the corresponding conjugate was labeled with 64Cu. Binding to TF and the stability was evaluated in vitro. TF targeting capability of the radiolabeled conjugate was tested in vivo by positron emission tomography (PET) imaging in pancreatic human xenograft cancer mouse models with various TF expressions. The conjugate showed good stability (>91% at 16 h), an immunoreactivity of 93.5%, and a mean tumor uptake of 2.1 ± 0.2%ID/g at 15 h post injection. In conclusion, FVIIai was radiolabeled with 64Cu in single well-defined position of the protein. This method can be utilized to prepare conjugates from serine proteases with the label at a specific position.

U2 - 10.1021/acs.bioconjchem.7b00649

DO - 10.1021/acs.bioconjchem.7b00649

M3 - Journal article

C2 - 29206443

AN - SCOPUS:85040732552

SN - 1043-1802

VL - 29

SP - 117

EP - 125

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 1