Fluorescence Imaging of the Lymph Node Uptake of Proteins in Mice after Subcutaneous Injection: Molecular Weight Dependence
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To use noninvasive fluorescence imaging to investigate the influence of molecular weight (MW) of proteins on the rate of loss from a subcutaneous (SC) injection site and subsequent uptake by the draining lymph nodes in mice.
Bevacizumab (149 kDa), bovine serum albumin (BSA, 66 kDa), ovalbumin (44.3 kDa) or VEGF-C156S (23 kDa), labeled with the near infrared dye IRDye 680, were injected SC into the front footpad of SKH-1 mice. Whole body non-invasive fluorescence imaging was performed to quantitate the fluorescence signal at the injection site and in axillary lymph nodes.
The half-life values, describing the times for 50% loss of proteins from the injection site, were 6.81 h for bevacizumab, 2.85 h for BSA, 1.57 h for ovalbumin and 0.31 h for VEGF-C156S. The corresponding axillary lymph node exposure, represented as the area of the % dose versus time curve, was 6.27, 5.13, 4.06 and 1.54% dose ∙ h, respectively.
Our results indicate that the rate of loss of proteins from a SC injection site is inversely related to MW of proteins, while lymph node exposure is proportionally related to the MW of proteins in a mouse model.
KEY WORDSfluorescence imaging lymphatic uptake molecular weight protein subcutaneous injection
bovine serum albumin
fraction of the dose recovered at the axillary lymph nodes
fraction of original signal remaining at the SC injection site
region of interest
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
vascular endothelial growth factor
ACKNOWLEDGMENTS & DISCLOSURES
This work is supported by a grant from the University at Buffalo Center for Protein Therapeutics to MEM. SGB was supported in part by a fellowship from Pfizer Global Research and Development. We acknowledge the valuable assistance from Dr. Rajiv Kumar, Lisa A. Vathy, Dr. Hong Ding, and Dr. Ken-Tye Yong from the Institute for Lasers, Photonics and Biophotonics, University at Buffalo.
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