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.
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