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Fluorescence Imaging of the Lymph Node Uptake of Proteins in Mice after Subcutaneous Injection: Molecular Weight Dependence

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ABSTRACT

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

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Abbreviations

BSA:

bovine serum albumin

FLN :

fraction of the dose recovered at the axillary lymph nodes

FSC :

fraction of original signal remaining at the SC injection site

IRDye:

infrared dye

LN:

lymph node

MW:

molecular weight

ROI:

region of interest

SC:

subcutaneous

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

VEGF:

vascular endothelial growth factor

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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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Author notes

  1. Suraj G. Bhansali

    Present address: Novartis Pharmaceuticals Corporation, Clinical PKPD, East Hanover, New Jersey, 07936, USA

Authors and Affiliations

  1. Department of Pharmaceutical Sciences School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, 517 Hochstetter Hall, Amherst, New York, 14260, USA

    Fang Wu, Suraj G. Bhansali & Marilyn E. Morris

  2. Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Amherst, New York, 14260, USA

    Wing Cheung Law, Earl J. Bergey & Paras N. Prasad

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  1. Fang Wu
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Correspondence to Marilyn E. Morris.

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Wu, F., Bhansali, S.G., Law, W.C. et al. Fluorescence Imaging of the Lymph Node Uptake of Proteins in Mice after Subcutaneous Injection: Molecular Weight Dependence. Pharm Res 29, 1843–1853 (2012). https://doi.org/10.1007/s11095-012-0708-6

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  • DOI: https://doi.org/10.1007/s11095-012-0708-6

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