Clinical Translation Brochure - LI-COR Bio Technical Resources ...

IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials.
Clinical
Translation
1
NEAR-INFRARED DETECTION
PROVEN PERFORMANCE
IN THE LAB
PROMISING POTENTIAL
FOR THE CLINIC

1
LI-COR ® Biosciences’ near-infrared dye, IRDye ® 800CW, is
manufactured under current Good Manufacturing Practices
(cGMP), has completed a toxicity study performed under GLP
(Good Laboratory Practice), and has Drug Master Files (DMF)
on record with U.S. and European regulatory authorities. With
this work complete, the dye is ready for conjugation to targeted
biomolecules for use in clinical trials for image-guided surgery.
Why Image-Guided Surgery?
Surgical oncology is the cornerstone of treatment for solid
tumors and a patient’s survival is directly related to the quality
of the surgical resection. 1-9 Inadequate surgical margins
represent a high risk for adverse clinical outcomes following
cancer surgery. Successful clinical trials of targeted nearinfrared
(NIR) contrast agents and appropriate camera systems
that provide real-time, image-assisted vision to the surgeon,
may lead to improved patient clinical outcomes.
Molecular Imaging and Cancer
The feasibility of using fluorescent-labeled molecularly
targeted agents to image tumors in real time was recently
demonstrated by van Dam et al., 10 in which a contrast agent
consisting of a folate ligand linked to fluorescein isothiocyanate
was used to target the folate receptor specifically expressed
in ovarian cancer. However, the tissue-scattering properties,
autofluorescence, and absorption of light in the 400- to 650-nm
References
IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials.
Prostate Cancer Surgical Margins
1 Emerson et al. J Clin Pathol. 2005; 58:1028-1032.
Esophageal Cancer Margins
2 Deeter et al. Arch Surg. 2009; 144(7):618-624.
Colorectal Cancer
3 Nagtegaal et al. J Clin Oncol. 2008; Vol 26, No 2.
Scan this QR code or visit www.licor.com/IRexcision to
see the video of this preclinical lymph node excision
from a mouse.
range where fluorescein absorbs and emits, makes the use
of NIR fluorophores more attractive due to minimization of
these interfering effects in this spectral region, especially for
deep-seated tumors.
10 Van Dam, et al. Nature Medicine 17, 1315-1319 (2011).
Breast Cancer
4 Jacobs. Ann Surg Oncol. 2008; 15:1271–2.
5 Kreike et al. Int J Radiat Oncol Biol Phys. 2008; 71:1014–21.
6 Rubio et al. Cir Esp. 2006; 79:3–9.
7 Miller et al. J Surg Oncol. 2004; 86:134–40.
8 Weng et al. Cancer. 2000; 88:1643–9.
9 Park et al. J Clin Oncol. 2000; 18:1668–75.
For a complete list of references related to the importance of positive surgical margins in cancer care, please visit www.licor.com/cancermargins.

IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials. 2
Are You Ready to Begin Translating Your Research?
Start with IRDye 800CW
When choosing the dye that you will need to begin
translating your research to the clinic, the following
points are important to consider:
Your dye must be manufactured under a cGMP process
Your cGMP dye must be available in quantities suitable
for extended stability and toxicity studies
A toxicity study of your dye must be completed under GLP
A DMF must be submitted to the appropriate
government agencies (i.e. United States Food and Drug
Administration or European Medicines Agency) to make
your application easier and faster
Your dye must be compatible with instruments that
have been cleared for clinical use
IRDye 800CW is ready for use
in your clinical trials
Current Collaborations
LI-COR ® Biosciences and Spectros Corporation
announced a non-exclusive licensing and
supply agreement in December 2009, to couple
IRDye 800CW to Spectros’ proprietary targeted
biomarkers, and to develop the targeted
fluorescent contrast agents for real-time imaging
and treatment of prostate and ovarian cancers in
patients. www.licor.com/SpectrosPR
LI-COR Biosciences has followed and completed these
key steps toward the potential translation of IRDye ®
800CW into clinical use:
IRDye 800CW is manufactured under a cGMP process
IRDye 800CW manufacturing has been scaled up in
quantities sufficient for preclinical and clinical studies
The toxicity study of IRDye 800CW in Sprague-Dawley ®
rats was reported in December 2010 in Molecular
Imaging and Biology Mol. Imaging Biol. 12:583-594 (2010).
A DMF for IRDye 800CW N-succidimidyl ester has
been filed with the United States Food and Drug
Administration in anticipation of clinical trials in the
United States
An Active Substance Master File (ASMF) for IRDye
800CW N-succidimidyl ester has been registered with
European Medicines Agency (EMA) in support of a
clinical trial
IRDye 800CW is compatible with instruments that
have been cleared for clinical use and that can detect
indocyanine green
Dutch Clinical Trial: NL37479.v042.11
Validation of Uptake of a VEGF-targeted Optical Fluorescent Imaging Tracer in
Surgical Specimens of Breast Cancer and Application of Pre- and Intra-operative
Human Molecular Fluorescence Imaging Techniques. A multicenter feasibility study.
Learn more at www.licor.com/translational
For updated information on these and other collaborations, please visit www.licor.com/collaborations.
University Medical Center Groningen is working
with LI-COR Biosciences and IRDye 800CW
to explore new tools for targeted fluorescent
imaging probes that will help address the
challenges facing oncology surgeons.

Extinction coefficient (hemoglobin)
3
Choosing Near-Infrared Detection
Optical imaging takes place optimally in the near-infrared
region due to low background. Tissue strongly absorbs and
scatters visible light. This limits the penetration of excitation
light and the detection of emitted fluorescence for detection.
At NIR wavelengths, tissue absorption is dramatically reduced
(Fig. 1) and light penetration is improved.
1000000
100000
10000
1000
IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials.
GFP
Cy3
Figure 1. Animal tissue absorbs visible light. Hemoglobin (Hb)
and other tissue components strongly absorb visible light. In the
NIR region, where infrared agents are detected, tissue absorbance
is dramatically reduced. Above 820 nm, light absorbance by water
begins to increase.
Imaging systems appropriate for investigational use with excitation and emission
wavelengths compatible with IRDye 800CW:
Carl Zeiss Meditec Clinical Solutions OPMI Pentaro C
with INFRARED 800
NOVADAQ ® SPY ®
Intuitive Surgical ® da Vinci Surgical System
O2View BV Artemis
Near-Infrared
Imaging Window
Cy5
IRDye 680RD
Wavelength (nm)
IRDye 800CW
Leica Microsystems FL800 for Integrated
Vascular Fluorescence
1.000
0.100
Hb
Water
100
0.010
400 500 600 700 800 900 1000
Extinction coefficient (water)
Tissue autofluorescence is significantly reduced in the NIR
range, so the desired signals are not obscured. Smaller doses
of optical agents can typically be used, and detection of the
early stages of disease may become possible.
Other approaches you may have considered are bioluminescent
imaging and fluorescent optical imaging with an NIR dye
called indocyanine green (ICG). Both of these approaches
have limitations for clinical translation. Bioluminescence
requires genetically engineered cell lines and transgenic
animals, so it does not have the potential to translate to clinical
applications. ICG is currently used in both preclinical and
clinical work; however, it is not available with a reactive group
for conjugation to a targeting moiety for molecular imaging.
Near-infrared imaging agents like LI-COR ® Biosciences’ IRDye ®
800CW, however, do have the potential for clinical translation.
IRDye 800CW is one of the only functionalized dyes that meets
the criteria for use in clinical trials. Laboratory studies have
shown that IRDye 800CW has several additional advantages
that include:
Brightness 20 to 50 times greater than ICG
Unlike ICG, its clearance route is through
the kidney rather than the liver, as shown in
laboratory studies
Fluoptics, Minatec-BHT Fluobeam ®
SurgOptix
Mizuho Medical Co. HyperEye Medical System
Karl Storz Endoscopes
Frangioni, FLARE ® System

in vitro
IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials. 4
in vivo
One Dye Simplifies Discovery
Start Your Preclinical Probe Development
with the Clinic in Mind
Starting your research with IRDye 800CW means you are one
step closer to the potential for clinical translation. By choosing
any of the other NIR dyes in LI-COR Biosciences’ portfolio of
preclinical products, you can have confidence that your research
will be successful from the initial Western blot to potential
clinical translation.
One dye can be used from the early stages of development,
and on to potential clinical translation. LI-COR can be your
solution for optical probe development and validation from
cells, to animals, to ex vivo validation.
How to Begin
Feasibility of Photoimmunotherapy in Tumor-bearing Mice
LI-COR offers ready-to-use NIR dyes and dye
labeling kits (see Table 1 on page 5) that have been
optimized for small animal imaging in both the 700
nm and 800 nm ranges. As with all LI-COR products,
these come with personalized technical support.
Choose an imager with the ultimate ease of use with
the freedom to work from the cell to the animal.
Utilize well-characterized controls – pre-optimized
optical probes work to confirm co-localization with
your probe.
Image organs and tissue sections ex vivo to validate
in vivo results
M. Mitsunaga et al. 2011.
Cancer cell–selective in vivo near-infrared photoimmunotherapy targeting specific membrane molecules
Nature Medicine 17: 1685–1691
Mitsunaga et al. developed a new type of cancer photoimmunotherapy
with a target-specific photosensitizer based on
IRDye ® 700DX conjugated to panitumumab and trastuzamab,
monoclonal antibodies targeting epidermal growth factor receptors
(EGFRs). Observed tumor shrinkage in mouse xenograph tumor
models expressing EGFRs after NIR irradiation. The antibody
whole organ tissue
IRDye 700DX conjugates were most effective when bound to the
cell membrane and produced no phototoxicity when not bound.
Further, because IRDye 700DX also emits a diagnostic fluorescence,
the therapeutic light could be targeted to tumor areas to minimize
exposure to nonrelevant tissues and to noninvasively monitor
therapeutic effects.

5
IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials.
Research Products to Complement
Clinical Translation
Dyes and Labeling Kits
IRDye ® infrared dye and protein labeling kits label antibodies and other proteins for applications such as In-Cell Western
assays, in vivo imaging, and whole organ or tissue section imaging (Table 1). These kits are convenient for researchers who
want to try the reagents before committing to large quantities. After the appropriate dyes and reagents are identified, affordable
scale-up quantities (50 mg) are available from LI-COR ® . IRDye research products are manufactured under non-cGMP and are
for research use in the laboratory only.
Table 1. IRDye infrared dyes and protein labeling kits available from LI-COR
Product Description P/N
IRDye 800CW NHS Ester Labels primary and secondary amines, such as lysine residues in proteins 929-70021
IRDye 800CW Maleimide Labels free sulfhydryl (—SH) groups, such as cysteine residues in proteins 929-80021
IRDye 800CW Carboxylate Non-reactive form of dye; used as control for dye retention 929-09406
IRDye 700DX NHS Ester Labels primary and secondary amines, such as lysine residues in proteins 929-70011
IRDye 680RD NHS Ester Labels primary and secondary amines, such as lysine residues in proteins 929-70051
IRDye 680RD Maleimide Labels free sulfhydryl (—SH) groups, such as cysteine residues in proteins 929-71051
IRDye 680RD Carboxylate Non-reactive form on dye; used as control for dye retention 929-08980
IRDye 800CW Protein Labeling Kit, High MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (45-200 kDa) 928-38040
IRDye 800CW Protein Labeling Kit, Low MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (15-45 kDa) 928-38042
IRDye 800CW Protein Labeling Kit, Microscale NHS ester reactive dye; for labeling of 3 x 100 µg protein (14-200 kDa) 928-38044
IRDye 700DX Protein Labeling Kit, High MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (45-200 kDa) 928-38046
IRDye 700DX Protein Labeling Kit, Low MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (15-45 kDa) 928-38048
IRDye 700DX Protein Labeling Kit, Microscale NHS ester reactive dye; for labeling of 3 x 100 µg protein (14-200 kDa) 928-38050
IRDye 680RD Protein Labeling Kit, High MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (45-200 kDa) 928-38072
IRDye 680RD Labeling Kit, Low MW NHS ester reactive dye; for labeling of 3 x 1 mg protein (15-45 kDa) 928-38074
IRDye 680RD Labeling Kit, Microscale NHS ester reactive dye; for labeling of 3 x 100 µg protein (14-200 kDa) 928-38076
Pearl ®
Impulse
Small Animal Imaging System
The Pearl Impulse Small Animal Imaging System offers in vivo molecular imaging with speed,
sensitivity, simplicity, and a sensible price. The Pearl Impulse incorporates near-infrared laser excitation
and a unique filtering design for exceptional performance. For researchers working toward the clinic,
the Pearl Impulse is designed to align with many available clinical fluorescent imaging systems.
Odyssey ®
CLx
Infrared Imaging System
The Odyssey CLx Infrared Imaging System accommodates a variety of
sample formats, including tissue sections, cell-based microplate assays,
and quantitative Western blots.

IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials. 6
Preclinical Applications of IRDye
Infrared Dye Optical Probes *
Lymph node metastasis in mouse with orthotopic
prostate tumor, imaged with IRDye 800CW EGF.
Arthritis imaging with IRDye 800CW 2-DG in DBA Collagen
antibody-induced arthritis model.
Lymph node imaging with IRDye 680RD HA, inset image
with white light.
*All images acquired using Pearl Impulse Small Animal Imaging System
Tumor vasculature imaged
with IRDye 800CW PEG.
Subcutaneous U87 tumor with
elevated integrin expression
detected on the mouse’s hip,
imaged with IRDye 680RD RGD.
www.licor.com/clinical
Micro-metastasis to kidney
(intraoperative imaging)
Subcutaneous A431 tumor with increased
glucose metabolism detected with IRDye
800CW 2-DG.
Visit our Clinical Translation homepage for a video about current research using nearinfrared
fluorescent imaging during surgery, as well as the latest in relevant news, information,
and publications.
Please contact us for more information about IRDye 800CW or the potential translation
of your research to the clinic.
Email Clinical.Translation@licor.com
Tel (402) 467-0904

U.S.
LI-COR ® Biosciences
4647 Superior Street
Lincoln, NE 68504
Phone: 402-467-0700
Phone: 800-645-4267
Fax: 402-467-0819
Email: biosales@licor.com
IRDye ® 800CW manufactured under cGMP is intended only for investigational use in clinical trials.
Locations WorldWide
LI-COR GmbH, Germany
Serving Europe, Africa, and the Middle East
LI-COR Biosciences GmbH
Siemensstraße 25A
D-61352 Bad Homburg
Germany
Phone: +49 (0) 6172 17 17 771
Fax: +49 (0) 6172 17 17 799
Email: bio-eu@licor.com
LI-COR Ltd., UK
Serving UK, Ireland, and Scandinavia
LI-COR Biosciences UK Ltd
St. John’s Innovation Centre
Cowley Road
Cambridge
CB4 0WS
United Kingdom
Phone: +44 (0) 1223 422104
Fax: +44 (0) 1223 422105
Email: bio-eu@licor.com
View a complete list of our international distributors at:
www.licor.com/bio/distributors
LI-COR is an ISO 9001 registered company. © 2012 LI-COR, Inc. Specifications
subject to change. LI-COR, Pearl, Odyssey, BrightSite, In-Cell Western, and IRDye are
trademarks or registered trademarks of LI-COR, Inc. in the United States and other
countries. All other trademarks belong to their respective owners. The Pearl Impulse
Imager, Odyssey Infrared Imaging System, and IRDye Infrared Dye technologies are
covered by U.S. Patents, foreign equivalents, and other patents pending.
980-13325 10/12
The LI-COR board of directors would like to take this opportunity to return thanks to
God for His merciful providence in allowing LI-COR to develop and commercialize
products, through the collective effort of dedicated employees, that enable the
examination of the wonders of His works.
“Trust in the LORD with all your heart and do not lean on your own understanding.
In all your ways acknowledge Him, and He will make your paths straight.”
—Proverbs 3:5,6