Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease

Hang T Ta, Sandeep Prabhu, Ephraem Leitner, Fu Jia, Dominik von Elverfeldt, Katherine E Jackson, Timo Heidt, Ashish Kumar Narayan Nair, Hannah Pearce, Constantin Von Zur Muhlen, Xiaowei Wang, Karlheinz Peter, Christoph Eugen Hagemeyer

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Antibody-targeted delivery of imaging agents can enhance the sensitivity and accuracy of current imaging techniques. Similarly, homing of effector cells to disease sites increases the efficacy of regenerative cell therapy while reducing the number of cells required. Currently, targeting can be achieved via chemical conjugation to specific antibodies, which typically results in the loss of antibody functionality and in severe cell damage. An ideal conjugation technique should ensure retention of antigen-binding activity and functionality of the targeted biological component. Objective: To develop a biochemically robust, highly reproducible, and site-specific coupling method using the Staphylococcus aureus sortase A enzyme for the conjugation of a single-chain antibody (scFv) to nanoparticles and cells for molecular imaging and cell homing in cardiovascular diseases. This scFv specifically binds to activated platelets, which play a pivotal role in thrombosis, atherosclerosis, and inflammation. Methods and Results: The conjugation procedure involves chemical and enzyme-mediated coupling steps. The scFv was successfully conjugated to iron oxide particles (contrast agents for magnetic resonance imaging) and to model cells. Conjugation efficiency ranged between 50 and 70 , and bioactivity of the scFv after coupling was preserved. The targeting of scFv-coupled cells and nanoparticles to activated platelets was strong and specific as demonstrated in in vitro static adhesion assays, in a flow chamber system, in mouse intravital microscopy, and in in vivo magnetic resonance imaging of mouse carotid arteries. Conclusions: This unique biotechnological approach provides a versatile and broadly applicable tool for procuring targeted regenerative cell therapy and targeted molecular imaging in cardiovascular and inflammatory diseases and beyond.
Original languageEnglish
Pages (from-to)365 - 373
Number of pages9
JournalCirculation Research
Volume109
Issue number4
DOIs
Publication statusPublished - 2011
Externally publishedYes

Cite this

Ta, Hang T ; Prabhu, Sandeep ; Leitner, Ephraem ; Jia, Fu ; von Elverfeldt, Dominik ; Jackson, Katherine E ; Heidt, Timo ; Nair, Ashish Kumar Narayan ; Pearce, Hannah ; Von Zur Muhlen, Constantin ; Wang, Xiaowei ; Peter, Karlheinz ; Hagemeyer, Christoph Eugen. / Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease. In: Circulation Research. 2011 ; Vol. 109, No. 4. pp. 365 - 373.
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title = "Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease",
abstract = "Antibody-targeted delivery of imaging agents can enhance the sensitivity and accuracy of current imaging techniques. Similarly, homing of effector cells to disease sites increases the efficacy of regenerative cell therapy while reducing the number of cells required. Currently, targeting can be achieved via chemical conjugation to specific antibodies, which typically results in the loss of antibody functionality and in severe cell damage. An ideal conjugation technique should ensure retention of antigen-binding activity and functionality of the targeted biological component. Objective: To develop a biochemically robust, highly reproducible, and site-specific coupling method using the Staphylococcus aureus sortase A enzyme for the conjugation of a single-chain antibody (scFv) to nanoparticles and cells for molecular imaging and cell homing in cardiovascular diseases. This scFv specifically binds to activated platelets, which play a pivotal role in thrombosis, atherosclerosis, and inflammation. Methods and Results: The conjugation procedure involves chemical and enzyme-mediated coupling steps. The scFv was successfully conjugated to iron oxide particles (contrast agents for magnetic resonance imaging) and to model cells. Conjugation efficiency ranged between 50 and 70 , and bioactivity of the scFv after coupling was preserved. The targeting of scFv-coupled cells and nanoparticles to activated platelets was strong and specific as demonstrated in in vitro static adhesion assays, in a flow chamber system, in mouse intravital microscopy, and in in vivo magnetic resonance imaging of mouse carotid arteries. Conclusions: This unique biotechnological approach provides a versatile and broadly applicable tool for procuring targeted regenerative cell therapy and targeted molecular imaging in cardiovascular and inflammatory diseases and beyond.",
author = "Ta, {Hang T} and Sandeep Prabhu and Ephraem Leitner and Fu Jia and {von Elverfeldt}, Dominik and Jackson, {Katherine E} and Timo Heidt and Nair, {Ashish Kumar Narayan} and Hannah Pearce and {Von Zur Muhlen}, Constantin and Xiaowei Wang and Karlheinz Peter and Hagemeyer, {Christoph Eugen}",
year = "2011",
doi = "10.1161/CIRCRESAHA.111.249375",
language = "English",
volume = "109",
pages = "365 -- 373",
journal = "Circulation Research",
issn = "0009-7330",
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Ta, HT, Prabhu, S, Leitner, E, Jia, F, von Elverfeldt, D, Jackson, KE, Heidt, T, Nair, AKN, Pearce, H, Von Zur Muhlen, C, Wang, X, Peter, K & Hagemeyer, CE 2011, 'Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease' Circulation Research, vol. 109, no. 4, pp. 365 - 373. https://doi.org/10.1161/CIRCRESAHA.111.249375

Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease. / Ta, Hang T; Prabhu, Sandeep; Leitner, Ephraem; Jia, Fu; von Elverfeldt, Dominik; Jackson, Katherine E; Heidt, Timo; Nair, Ashish Kumar Narayan; Pearce, Hannah; Von Zur Muhlen, Constantin; Wang, Xiaowei; Peter, Karlheinz; Hagemeyer, Christoph Eugen.

In: Circulation Research, Vol. 109, No. 4, 2011, p. 365 - 373.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Enzymatic single-chain antibody tagging: A universal approach to targeted molecular imaging and cell homing in cardiovascular disease

AU - Ta, Hang T

AU - Prabhu, Sandeep

AU - Leitner, Ephraem

AU - Jia, Fu

AU - von Elverfeldt, Dominik

AU - Jackson, Katherine E

AU - Heidt, Timo

AU - Nair, Ashish Kumar Narayan

AU - Pearce, Hannah

AU - Von Zur Muhlen, Constantin

AU - Wang, Xiaowei

AU - Peter, Karlheinz

AU - Hagemeyer, Christoph Eugen

PY - 2011

Y1 - 2011

N2 - Antibody-targeted delivery of imaging agents can enhance the sensitivity and accuracy of current imaging techniques. Similarly, homing of effector cells to disease sites increases the efficacy of regenerative cell therapy while reducing the number of cells required. Currently, targeting can be achieved via chemical conjugation to specific antibodies, which typically results in the loss of antibody functionality and in severe cell damage. An ideal conjugation technique should ensure retention of antigen-binding activity and functionality of the targeted biological component. Objective: To develop a biochemically robust, highly reproducible, and site-specific coupling method using the Staphylococcus aureus sortase A enzyme for the conjugation of a single-chain antibody (scFv) to nanoparticles and cells for molecular imaging and cell homing in cardiovascular diseases. This scFv specifically binds to activated platelets, which play a pivotal role in thrombosis, atherosclerosis, and inflammation. Methods and Results: The conjugation procedure involves chemical and enzyme-mediated coupling steps. The scFv was successfully conjugated to iron oxide particles (contrast agents for magnetic resonance imaging) and to model cells. Conjugation efficiency ranged between 50 and 70 , and bioactivity of the scFv after coupling was preserved. The targeting of scFv-coupled cells and nanoparticles to activated platelets was strong and specific as demonstrated in in vitro static adhesion assays, in a flow chamber system, in mouse intravital microscopy, and in in vivo magnetic resonance imaging of mouse carotid arteries. Conclusions: This unique biotechnological approach provides a versatile and broadly applicable tool for procuring targeted regenerative cell therapy and targeted molecular imaging in cardiovascular and inflammatory diseases and beyond.

AB - Antibody-targeted delivery of imaging agents can enhance the sensitivity and accuracy of current imaging techniques. Similarly, homing of effector cells to disease sites increases the efficacy of regenerative cell therapy while reducing the number of cells required. Currently, targeting can be achieved via chemical conjugation to specific antibodies, which typically results in the loss of antibody functionality and in severe cell damage. An ideal conjugation technique should ensure retention of antigen-binding activity and functionality of the targeted biological component. Objective: To develop a biochemically robust, highly reproducible, and site-specific coupling method using the Staphylococcus aureus sortase A enzyme for the conjugation of a single-chain antibody (scFv) to nanoparticles and cells for molecular imaging and cell homing in cardiovascular diseases. This scFv specifically binds to activated platelets, which play a pivotal role in thrombosis, atherosclerosis, and inflammation. Methods and Results: The conjugation procedure involves chemical and enzyme-mediated coupling steps. The scFv was successfully conjugated to iron oxide particles (contrast agents for magnetic resonance imaging) and to model cells. Conjugation efficiency ranged between 50 and 70 , and bioactivity of the scFv after coupling was preserved. The targeting of scFv-coupled cells and nanoparticles to activated platelets was strong and specific as demonstrated in in vitro static adhesion assays, in a flow chamber system, in mouse intravital microscopy, and in in vivo magnetic resonance imaging of mouse carotid arteries. Conclusions: This unique biotechnological approach provides a versatile and broadly applicable tool for procuring targeted regenerative cell therapy and targeted molecular imaging in cardiovascular and inflammatory diseases and beyond.

UR - http://circres.ahajournals.org/content/109/4/365

U2 - 10.1161/CIRCRESAHA.111.249375

DO - 10.1161/CIRCRESAHA.111.249375

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VL - 109

SP - 365

EP - 373

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

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