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Pharmaceutical Research

, Volume 29, Issue 7, pp 1843–1853 | Cite as

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

  • Fang Wu
  • Suraj G. Bhansali
  • Wing Cheung Law
  • Earl J. Bergey
  • Paras N. Prasad
  • Marilyn E. MorrisEmail author
Research Paper

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.

KEY WORDS

fluorescence imaging lymphatic uptake molecular weight protein subcutaneous injection 

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

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Fang Wu
    • 1
  • Suraj G. Bhansali
    • 1
    • 2
  • Wing Cheung Law
    • 3
  • Earl J. Bergey
    • 3
  • Paras N. Prasad
    • 3
  • Marilyn E. Morris
    • 1
    Email author
  1. 1.Department of Pharmaceutical Sciences School of Pharmacy and Pharmaceutical SciencesUniversity at Buffalo, State University of New YorkAmherstUSA
  2. 2.Novartis Pharmaceuticals Corporation, Clinical PKPDEast HanoverUSA
  3. 3.Institute for Lasers, Photonics and BiophotonicsUniversity at Buffalo, State University of New YorkAmherstUSA

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