High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting

Christopher Ullman, Pascale Mathonet, Arkadiusz Oleksy, Agata Diamamdakis, Licia Tomei, Anna Demartis, Maria Chiara Nardi, Sonia Sambucini, Antonino Missineo, Karen Maria Alt, Christoph Eugen Hagemeyer, Matthew Harris, Amos Hedt, Roland Weis, Kurt Ronald Gehlsen

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Abdurins are a novel antibody-like scaffold derived from the engineering of a single isolated CH2 domain of human IgG. Previous studies established the prolonged serum half-life of Abdurins, the result of a retained FcRn binding motif. Here we present data on the construction of large, diverse, phage-display and cell-free DNA display libraries and the isolation of high affinity binders to the cancer target, membrane-bound ephrin receptor tyrosine kinase class A2 (EphA2). Antigen binding regions were created by designing combinatorial libraries into the structural loops and Abdurins were selected using phage display methods. Initial binders were reformatted into new maturation libraries and low nanomolar binders were isolated using cell-free DNA display, CIS display. Further characterization confirmed binding of the Abdurins to both human and murine EphA2 proteins and exclusively to cell lines that expressed EphA2, followed by rapid internalization. Two different EphA2 binders were labeled with 64Cu, using a bifunctional MeCOSar chelator, and administered to mice bearing tumors from transplanted human prostate cancer cells, followed by PET/CT imaging. The anti-EphA2 Abdurins localized in the tumors as early as 4 hours after injection and continued to accumulate up to 48 hours when the imaging was completed. These data demonstrate the ability to isolate high affinity binders from the engineered Abdurin scaffold, which retain a long serum half-life, and specifically target tumors in a xenograft model.
Original languageEnglish
Article numbere0135278
Number of pages25
JournalPLoS ONE
Volume10
Issue number8
DOIs
Publication statusPublished - 2015
Externally publishedYes

Cite this

Ullman, C., Mathonet, P., Oleksy, A., Diamamdakis, A., Tomei, L., Demartis, A., ... Gehlsen, K. R. (2015). High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting. PLoS ONE, 10(8), [e0135278]. https://doi.org/10.1371/journal.pone.0135278
Ullman, Christopher ; Mathonet, Pascale ; Oleksy, Arkadiusz ; Diamamdakis, Agata ; Tomei, Licia ; Demartis, Anna ; Nardi, Maria Chiara ; Sambucini, Sonia ; Missineo, Antonino ; Alt, Karen Maria ; Hagemeyer, Christoph Eugen ; Harris, Matthew ; Hedt, Amos ; Weis, Roland ; Gehlsen, Kurt Ronald. / High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting. In: PLoS ONE. 2015 ; Vol. 10, No. 8.
@article{becb525d5fcc452ba2adcfa207d358c0,
title = "High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting",
abstract = "Abdurins are a novel antibody-like scaffold derived from the engineering of a single isolated CH2 domain of human IgG. Previous studies established the prolonged serum half-life of Abdurins, the result of a retained FcRn binding motif. Here we present data on the construction of large, diverse, phage-display and cell-free DNA display libraries and the isolation of high affinity binders to the cancer target, membrane-bound ephrin receptor tyrosine kinase class A2 (EphA2). Antigen binding regions were created by designing combinatorial libraries into the structural loops and Abdurins were selected using phage display methods. Initial binders were reformatted into new maturation libraries and low nanomolar binders were isolated using cell-free DNA display, CIS display. Further characterization confirmed binding of the Abdurins to both human and murine EphA2 proteins and exclusively to cell lines that expressed EphA2, followed by rapid internalization. Two different EphA2 binders were labeled with 64Cu, using a bifunctional MeCOSar chelator, and administered to mice bearing tumors from transplanted human prostate cancer cells, followed by PET/CT imaging. The anti-EphA2 Abdurins localized in the tumors as early as 4 hours after injection and continued to accumulate up to 48 hours when the imaging was completed. These data demonstrate the ability to isolate high affinity binders from the engineered Abdurin scaffold, which retain a long serum half-life, and specifically target tumors in a xenograft model.",
author = "Christopher Ullman and Pascale Mathonet and Arkadiusz Oleksy and Agata Diamamdakis and Licia Tomei and Anna Demartis and Nardi, {Maria Chiara} and Sonia Sambucini and Antonino Missineo and Alt, {Karen Maria} and Hagemeyer, {Christoph Eugen} and Matthew Harris and Amos Hedt and Roland Weis and Gehlsen, {Kurt Ronald}",
year = "2015",
doi = "10.1371/journal.pone.0135278",
language = "English",
volume = "10",
journal = "PLoS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "8",

}

Ullman, C, Mathonet, P, Oleksy, A, Diamamdakis, A, Tomei, L, Demartis, A, Nardi, MC, Sambucini, S, Missineo, A, Alt, KM, Hagemeyer, CE, Harris, M, Hedt, A, Weis, R & Gehlsen, KR 2015, 'High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting', PLoS ONE, vol. 10, no. 8, e0135278. https://doi.org/10.1371/journal.pone.0135278

High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting. / Ullman, Christopher; Mathonet, Pascale; Oleksy, Arkadiusz; Diamamdakis, Agata; Tomei, Licia; Demartis, Anna; Nardi, Maria Chiara; Sambucini, Sonia; Missineo, Antonino; Alt, Karen Maria; Hagemeyer, Christoph Eugen; Harris, Matthew; Hedt, Amos; Weis, Roland; Gehlsen, Kurt Ronald.

In: PLoS ONE, Vol. 10, No. 8, e0135278, 2015.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - High affinity binders to EphA2 isolated from Abdurin Scaffold libraries; Characterization, binding and tumor targeting

AU - Ullman, Christopher

AU - Mathonet, Pascale

AU - Oleksy, Arkadiusz

AU - Diamamdakis, Agata

AU - Tomei, Licia

AU - Demartis, Anna

AU - Nardi, Maria Chiara

AU - Sambucini, Sonia

AU - Missineo, Antonino

AU - Alt, Karen Maria

AU - Hagemeyer, Christoph Eugen

AU - Harris, Matthew

AU - Hedt, Amos

AU - Weis, Roland

AU - Gehlsen, Kurt Ronald

PY - 2015

Y1 - 2015

N2 - Abdurins are a novel antibody-like scaffold derived from the engineering of a single isolated CH2 domain of human IgG. Previous studies established the prolonged serum half-life of Abdurins, the result of a retained FcRn binding motif. Here we present data on the construction of large, diverse, phage-display and cell-free DNA display libraries and the isolation of high affinity binders to the cancer target, membrane-bound ephrin receptor tyrosine kinase class A2 (EphA2). Antigen binding regions were created by designing combinatorial libraries into the structural loops and Abdurins were selected using phage display methods. Initial binders were reformatted into new maturation libraries and low nanomolar binders were isolated using cell-free DNA display, CIS display. Further characterization confirmed binding of the Abdurins to both human and murine EphA2 proteins and exclusively to cell lines that expressed EphA2, followed by rapid internalization. Two different EphA2 binders were labeled with 64Cu, using a bifunctional MeCOSar chelator, and administered to mice bearing tumors from transplanted human prostate cancer cells, followed by PET/CT imaging. The anti-EphA2 Abdurins localized in the tumors as early as 4 hours after injection and continued to accumulate up to 48 hours when the imaging was completed. These data demonstrate the ability to isolate high affinity binders from the engineered Abdurin scaffold, which retain a long serum half-life, and specifically target tumors in a xenograft model.

AB - Abdurins are a novel antibody-like scaffold derived from the engineering of a single isolated CH2 domain of human IgG. Previous studies established the prolonged serum half-life of Abdurins, the result of a retained FcRn binding motif. Here we present data on the construction of large, diverse, phage-display and cell-free DNA display libraries and the isolation of high affinity binders to the cancer target, membrane-bound ephrin receptor tyrosine kinase class A2 (EphA2). Antigen binding regions were created by designing combinatorial libraries into the structural loops and Abdurins were selected using phage display methods. Initial binders were reformatted into new maturation libraries and low nanomolar binders were isolated using cell-free DNA display, CIS display. Further characterization confirmed binding of the Abdurins to both human and murine EphA2 proteins and exclusively to cell lines that expressed EphA2, followed by rapid internalization. Two different EphA2 binders were labeled with 64Cu, using a bifunctional MeCOSar chelator, and administered to mice bearing tumors from transplanted human prostate cancer cells, followed by PET/CT imaging. The anti-EphA2 Abdurins localized in the tumors as early as 4 hours after injection and continued to accumulate up to 48 hours when the imaging was completed. These data demonstrate the ability to isolate high affinity binders from the engineered Abdurin scaffold, which retain a long serum half-life, and specifically target tumors in a xenograft model.

UR - http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0135278

U2 - 10.1371/journal.pone.0135278

DO - 10.1371/journal.pone.0135278

M3 - Article

VL - 10

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 8

M1 - e0135278

ER -