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Exp Biol Med (Maywood). 2014 Jul;239(7):842-852. Epub 2014 May 28.

Evaluation of antibody-chemokine fusion proteins for tumor-targeting applications.

Author information

1
Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology Zurich (ETH Zürich), CH-8093 Zürich, Switzerland.
2
Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology Zurich (ETH Zürich), CH-8093 Zürich, Switzerland neri@pharma.ethz.ch.

Abstract

There is an increasing biotechnological interest in the 'arming' of therapeutic antibodies with bioactive payloads. While many antibody-cytokine fusion proteins have been extensively investigated in preclinical and clinical studies, there are only few reports related to antibody-chemokine fusion proteins ('immunochemokines'). Here, we describe the cloning, expression, and characterization of 10 immunochemokines based on the monoclonal antibody F8, specific to the alternatively spliced extra domain A (EDA) of fibronectin, a marker of angiogenesis. Among the 10 murine chemokines tested in our study, only CCL19, CCL20, CCL21, and CXCL10 could be expressed and isolated at acceptable purity levels as F8-based fusion proteins. The immunochemokines retained the binding characteristics of the parental antibody, but could not be characterized by gel-filtration analysis, an analytical limitation which had previously been observed in our laboratory for the unconjugated chemokines. When radioiodinated preparations of CCL19-F8, CCL20-F8, CCL21-F8, and CXCL10-F8 were tested in quantitative biodistribution studies in tumor-bearing mice, the four fusion proteins failed to preferentially accumulate at the tumor site, while the unconjugated parental antibody displayed a tumor:blood ratio >20:1, 24 h after intravenous (i.v.) administration. The tumor-targeting ability of CCL19-F8 could be rescued only in part by preadministration of unlabeled CCL19-F8, indicating that a chemokine trapping mechanism may hinder pharmacodelivery strategies. While this article highlights expression, analytical, and biodistribution challenges associated with the antibody-based in vivo delivery of chemokines at sites of disease, it provides the first comprehensive report in this field and may facilitate future studies with immunochemokines.

KEYWORDS:

Immunochemokine; antibody–chemokine fusion protein; armed antibody; chemokine; oncofetal fibronectin; vascular targeting

PMID:
24872435
DOI:
10.1177/1535370214536667

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