in vivo imaging dye & probes - akina, inc · in vivo imaging dye & probes ... jae hyung...
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In vivo imaging dye & probes
Near-Infrared Fluorescent Dyes
Description
The Flamma® NIR Flours series from BioActs is a Near-Infrared fluorescent dye product for animal
imaging. BioActs products have high fluorescence intensity, high water solubility and low toxicity. The
NIR Fluorescent Dyes products from BioActs offers fluorescence wavelength ranges from 600 to 900 nm,
and small amount of interference by biomolecular autofluorescence occurs. With the effect of these
long wavelengths, it can be used as an effective imaging product for In-vivo experiments.
Flamma® Fluors Absorbance Spectra
Flamma® Fluors Emission Spectra
[Figure 1] Various absorption and fluorescence wavelengths of BioActs NIR products
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BioActs' NIR Dye contains bioreactor such as NHS ester, Maleimide, Isothiocyanate, etc., so you
can select the proper reactor you want and conduct effective experiments.
[Figure 2] Selection of reactors for binding of biomaterials with amines
[Figure 3] Selection of reactors for binding of biomolecules to Thiol
Product List
Cat. No. Product name Emission
color
ExMax
(nm)
EmMax
(nm)
Common
filter set Excitation laser line Size
PWC1201 Flamma® 648 Carboxylic acid ● Red
648 663
Cy®5 594, 633 nm 5mg, 25mg
PWC1501 Flamma® 675 Carboxylic acid ● Far red
675 691
Cy®5.5 633, 680 nm 5mg, 25mg
PWC1308 Flamma® 749 Carboxylic acid ● NIR
749 774
Cy®7 680 nm 5mg, 25mg
PWC1603 Flamma® 774 Carboxylic acid ● NIR
774 806
Cy®7.5 785 nm 5mg, 25mg
POC1616 ICG Carboxylic acid ● NIR
785 821
Cy®7.5 785 nm 5mg, 25mg
PWS1215 Flamma® 648 NHS ester ● Red
648 663
Cy®5 594, 633 nm 1mg, 5mg, 25mg
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PWS1515 Flamma® 675 NHS ester ● Far red
675 691
Cy®5.5 633, 680 nm 1mg, 5mg, 25mg
PWS1301 Flamma® 749 NHS ester ● NIR
749 774
Cy®7 680 nm 1mg, 5mg, 25mg
PWS1603 Flamma® 774 NHS ester ● NIR
774 806
Cy®7.5 785 nm 1mg, 5mg, 25mg
POS1604 ICG NHS ester ● NIR
785 821
Cy®7.5 785 nm 1mg, 5mg, 25mg
PWSN1215 Flamma® 648 Sulfo-NHS ester ● Red
648 663
Cy®5 594, 633 nm 1mg, 5mg, 25mg
PWSN1515 Flamma® 675 Sulfo-NHS ester ● Far red
675 691
Cy®5.5 633, 680 nm 1mg, 5mg, 25mg
PWSN1301 Flamma® 749 Sulfo-NHS ester ● NIR
749 774
Cy®7 680 nm 1mg, 5mg, 25mg
PWSN1603 Flamma® 774 Sulfo-NHS ester ● NIR
774 806
Cy®7.5 785 nm 1mg, 5mg, 25mg
POSN1604 ICG Sulfo-NHS ester ● NIR
785 821
Cy®7.5 785 nm 1mg, 5mg, 25mg
PWA1215 Flamma® 648 Vinylsulfone ● Red
648 663
Cy®5 594, 633 nm 1mg, 5mg, 25mg
PWA1515 Flamma® 675 Vinylsulfone ● Far red
675 691
Cy®5.5 633, 680 nm 1mg, 5mg, 25mg
PWA1603 Flamma® 774 Vinylsulfone ● NIR
774 806
Cy®7.5 785 nm 1mg, 5mg, 25mg
POA1616 ICG Vinylsulfone ● NIR
785 821
Cy®7.5 785 nm 1mg, 5mg, 25mg
KWI1215 Flamma® 648 Isothiocyanate ● Red
648 663
Cy®5 594, 633 nm 1mg, 5mg, 25mg
KWI1515 Flamma® 675 Isothiocyanate ● Far red
675 691
Cy®5.5 633, 680 nm 1mg, 5mg, 25mg
PWI1603 Flamma® 774 Isothiocyanate ● NIR
774 806
Cy®7.5 785 nm 1mg, 5mg, 25mg
POI1616 ICG Isothiocyanate ● NIR
785 821
Cy®7.5 785 nm 1mg, 5mg, 25mg
In vivo behavior experiment of NIR Dye near-infrared products
- The experimental procedure for in-vivo imaging of the product is as follows.
- Preparation: Balb/c nude male 5weeks old (We carry out the experiment at week 6 after 1
week stabilization period.
- Animal experiments: Dilute each fluorescent substance to a concentration of 0.03 mg / ml. is
Inject 100 μl into the prepared animal by intravenous injection. Observed Fluorescent images
at 1 minute, 30 minutes, 1 hour, 2 hours, 3 hours, and 1 day intervals. (Observation
equipment: IVIS)
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In-vivo Experiment result
[Figure 4] After 1 day of Flamma® 749 Carboxylic acid injection, elimination from the body was confirmed
[Figure 5] After 1 day of Flamma® 774 Carboxylic acid injection, elimination from the body was confirmed
[Figure 6] After 1 day of ICG Carboxylic acid injection, elimination from the body was confirmed)
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References
- Paclitaxel loaded hyaluronic acid nanoparticles for targeted cancer therapy: In vitro and in vivo analysis
(Reju G. Thomas, , MyeongJu Moon, SeJy Lee, Yong Yeon Jeong, International Journal of Biological
Macromolecules, 2015, Volume 72, Pages 510–518)
- Photo-crosslinked hyaluronic acid nanoparticles with improved stability for in vivo tumor-targeted drug
delivery (Hong Yeol Yoona, Heebeom Koo, Ki Young Choi, Ick Chan Kwon, Kuiwon Choi, Jae Hyung
Park, Biomaterials, 2013, Volume 34, Issue 21, Pages 5273–5280)
- Co-delivery of VEGF and Bcl-2 dual-targeted siRNA polymer using a single nanoparticle for synergistic
anti-cancer effects in vivo (So Jin Lee, Simmyung Yook, Ji Young Yhee, Hong Yeol Yoon, Myung-Goo
Kim, Sook Hee Ku, Sun Hwa Kim, Jae Hyung Park, Ji Hoon Jeong, Ick Chan Kwon, Seulki Lee, Hyukjin
Lee, Kwangmeyung Kim, Journal of Controlled Release, 2015, Volume 220, Part B, Pages 631–641)
- Effectiveness of Losartan-Loaded Hyaluronic Acid (HA) Micelles for the Reduction of Advanced Hepatic
Fibrosis in C3H/HeN Mice Model (Reju George Thomas, Myeong Ju Moon, Jo Heon Kim, Jae Hyuk Lee,
Yong Yeon Jeong, PLoS ONE, 2015, 10(12): e0145512.)
- Notch1 targeting siRNA delivery nanoparticles for rheumatoid arthritis therapy (Min Ju Kim, Jong-Sung
Park, So Jin Lee, Jiyeon Jang, Jin Su Park, Seung Hyun Back, Gahee Bahn, Jae Hyung Park, Young Mo
Kang, Sun Hwa Kim, Ick Chan Kwon, Dong-Gyu Jo, Kwangmeyung Kim, Journal of Controlled Release,
2015, Volume 216, Pages 140–148)
- Cancer-targeted MDR-1 siRNA delivery using self-cross-linked glycol chitosan nanoparticles to overcome
drug resistance (Ji Young Yhee, Seungyong Song, So Jin Lee, Sung-Gurl Park, Ki-Suk Kim, Myung Goo
Kim, Sejin Son, Heebeom Koo, Ick Chan Kwon, Ji Hoon Jeong, Seo Young Jeong, Sun Hwa Kim,
Kwangmeyung Kim, Journal of Controlled Release, 2015, Volume 198, Pages 1–9)
- A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT)
using Tat peptide-conjugated IgM (Kyung oh Jung, Hyewon Youn, Seung Hoo Kim, Young-Hwa Kim,
Keon Wook Kang, June-Key Chung, Biochemical and Biophysical Research Communications, 2016,
Volume 477, Issue 3, Pages 483–489)
- Controlled Release of Hepatocyte Growth Factor from MPEG-b-(PCL-ran-PLLA) Diblock Copolymer for
Improved Vocal Fold Regeneration (Jae Won Choi, Yeon Soo Kim, Ju Kyeong Park, Eun Hye Song, Ji
Hoon Park, Moon Suk Kim, Yoo Seob Shin, Chul-Ho Kim, Macromolecular Bioscience, 2016,
DOI: 10.1002/mabi.201600163)
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AngioFlamma® series
Description
Integrin is a heterodimeric cell-surface receptor that binds to an arginine-glycine-aspartate (RGD)
sequence and mediates adhesion and fixation between cells and substrates outside the cell. Integrin is
also associated with cell signaling and gene expression associated with cell growth, migration and survival.
Integrin is used as a marker in disease studies because it is involved in blood clotting, inflammatory
reactions and cancer manifestations.
BioActs' AngioFlamma® products are based on RGD peptides and are fluorescent probe materials with
Flamma® Fluors fluorescence products that are used in vivo inflammatory reactions, detection of tumor
and angiogenesis
Product List
Cat. No. Product name Emission
color
ExMax
(nm) EmMax
(nm)
Common
filter set
Excitation
laser line Size
ARW1025 AngioFlamma® FAM ● Green 492 519 FITC 488 nm 0.5mg, 1mg, 5mg
ARW1011 AngioFlamma® 552 ● Yellow 550 565 TRITC 488, 532 nm 0.5mg, 1mg, 5mg
ARR1001 AngioFlamma® TAMRA ● Orange 543 575 TRITC 488, 532 nm 0.5mg, 1mg, 5mg
ARW1028 AngioFlamma® 560 ● Orange 560 589 TRITC 488, 532 nm 0.5mg, 1mg, 5mg
ARW1215 AngioFlamma® 648 ● Far red 648 663 Cy®5 594, 633 nm 0.5mg, 1mg, 5mg
ARW1501 AngioFlamma® 675 ● NIR 675 691 Cy®5.5 633, 680 nm 0.5mg, 1mg, 5mg
ARW1301 AngioFlamma® 749 ● NIR 749 774 Cy®7 680 nm 0.5mg, 1mg, 5mg
ARW1601 AngioFlamma® 774 ● NIR 774 806 Cy®7.5 785 nm 0.5mg, 1mg, 5mg
ARO1601 AngioFlamma® ICG ● NIR 785 821 Cy®7.5 785 nm 0.5mg, 1mg, 5mg
In-vivo Imaging Protocol
- The experiment procedure for in-vivo imaging of the product proceeds as follows.
- PREPARING XENOGRAFT TUMOR MODEL FOR IMAGING
1. MDA-MB-231 human breast cancer cells (1.0 x 107) should be injected subcutaneously on
the female nude mouse shoulder.
2. After the tumor cells have grown to about 50-80 mm3, administer the drug.
- INJECTION and FLUORESCENCE IMAGING
1. Fluorescence imaging was performed up to 24 hours after injection using the eXplore
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Optix system (ART Advanced Research Technologies Inc., Montreal, Canada) and
Flamma® 675 products, excitation (λex = 675 nm) and emission (λem = 698 nm) filter
sets.
2. Prepared mice were injected intravenously with 100 μg / 200 μl of AngioFlamma® 675
product and images were taken at set times.
In-vivo Imaging Result
[Figure 7] Breast cancer cell imaging using AngioFlamma® 675
References
- Zwitterionic Chitosan–Polyamidoamine Dendrimer Complex Nanoparticles as a pH-Sensitive Drug
Carrier (Karen C. Liu, Yoon Yeo, Mol. Pharmaceutics, 2013, 10 (5), pp 1695–1704)
10min 3h 6h 9h 12h 24h
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ApoFlamma® series
Description
The ApoFlamma® series is a family of fluorescent probe products for detecting apoptotic cells,
allowing the observation of cells through fluorescence imaging by binding to apoptotic cells. Among the
ApoFlamma® series, the ApoFlamma® PS products allows fluorescence imaging to be observed by
specifically binding to the phosphatidylserine exposed to the outer membrane of the apoptotic cell. The
ApoFlamma® H product allows fluorescent imaging to be observed by specifically binding to Histone 1
released to the outside by the apoptotic process. Both product lines can be observed by in vivo
experiments. In particular, near-infrared fluorescence products can be used without being affected by the
autofluorescence of animal.
[Figure 8] Imaging using ApoFlamma® PS 648 of the Etoposide-treated Hela cell
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[Figure 9] Imaging with ApoFlamma PS 675 of B16F10 cells treated with Etoposide
Product List
Cat. No. Product name Emission
color
ExMax
(nm) EmMax
(nm)
Common
filter set
Excitation
laser line Size
APW1025 ApoFlamma® PS FAM ● Green 492 519 FITC 488 nm 10 doses, 50 doses, 200 doses
APP1001 ApoFlamma® PS TAMRA ● Orange 543 575 TRITC 488, 532 nm 10 doses, 50 doses, 200 doses
APW1011 ApoFlamma® PS 552 ● Yellow 550 565 TRITC 488, 532 nm 10 doses, 50 doses, 200 doses
APW1215 ApoFlamma® PS 648 ● Red 648 663 Cy®5 594, 633 nm 10 doses, 50 doses, 200 doses
APW1501 ApoFlamma® PS 675 ● Far red 675 698 Cy®5.5 633, 680 nm 10 doses, 50 doses, 200 doses
APW1301 ApoFlamma® PS 749 ● NIR 750 782 Cy®7 680 nm 10 doses, 50 doses, 200 doses
APW1601 ApoFlamma® PS 774 ● NIR 777 802 Cy®7.5 785 nm 10 doses, 50 doses, 200 doses
APO1601 ApoFlamma® PS ICG ● NIR 785 821 Cy®7.5 785 nm 10 doses, 50 doses, 200 doses
AHW1025 ApoFlamma® H FAM ● Green 492 519 FITC 488 nm 10 doses, 50 doses, 200 doses
AHH1001 ApoFlamma® H TAMRA ● Orange 543 575 TRITC 488, 532 nm 10 doses, 50 doses, 200 doses
AHW1011 ApoFlamma® H 552 ● Yellow 550 565 TRITC 488, 532 nm 10 doses, 50 doses, 200 doses
AHW1215 ApoFlamma® H 648 ● Red 648 663 Cy®5 594, 633 nm 10 doses, 50 doses, 200 doses
AHW1501 ApoFlamma® H 675 ● Far red 675 698 Cy®5.5 633, 680 nm 10 doses, 50 doses, 200 doses
AHW1301 ApoFlamma® H 749 ● NIR 750 782 Cy®7 680 nm 10 doses, 50 doses, 200 doses
AHW1601 ApoFlamma® H 774 ● NIR 777 802 Cy®7.5 785 nm 10 doses, 50 doses, 200 doses
AHO1601 ApoFlamma® H ICG ● NIR 785 821 Cy®7.5 785 nm 10 doses, 50 doses, 200 doses
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In-vivo Imaging Protocol
- The experiment procedure for in-vivo imaging of the product proceeds as follows.
- Six-week-old Balb/c female nude mice were injected subcutaneously with 1 x 108 MDA231
tumor cells suspended in RPMI medium containing 10% FBS in the right femur.
- When the tumor size of the mice was grown to 1 cm, injected intravenously with 100 μL of
ApoFlamma® (50 μM each) via tail vein after isoflurane anesthesia.
- 30 minutes after the injection of ApoFlamma®, the tumor was observed via the eXplorer
Optix (GE healthcare, USA) imaging equipment.
In-vivo Imaging Result
[Figure 10] Imaging Apoptosis process of drug-injected tumor cells using ApoFlamma® H 774
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[Figure 11] Imaging Apoptosis process of rheumatoid using ApoFlamma® product
[Figure 12] Imaging images of protective drugs using ApoFlamma® H 774 with Parkinson's disease model
induced by injection of MPTP drug.
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[Figure 13] Apoptotic cells by camptothecin imaging with ApoFlamma® PS FAM
References
- Monitoring the correlation between I-uptake and apoptosis using Apoptosis-targeting peptide-1 (ApoPep-
1) (Kyung Oh Jung, Seung Hoo Kim, Hyewon Youn, Keon Kang, Dong Soo Lee, June-Key Chung, J Nucl
Med, 2011, vol. 52, no. supplement 1, 1761)
- Relationship between Apoptosis Imaging and Radioiodine Therapy in Tumor Cells with Different Sodium
Iodide Symporter Gene Expression (Kyung Oh Jung, Hyewon Youn, Young-Hwa Kim, Seunghoo Kim,
Juri Na, Yong-il Kim, Jin Woo Park, Keon Wook Kang, Dong Soo Lee, June-Key Chung, Molecular
Imaging, 2014: pp 1–9)
- Sodium [18F]Fluoride PET/CT in Myocardial Infarction (Jeong Hee Han, Sue Yeon Lim, Min Su Lee,
Won Woo Lee, Molecular Imaging and Biology, 2015, Volume 17, Issue 2, pp 214–221)
- A novel method to detect articular chondrocyte death during early stages of osteoarthritis using a non-
invasive ApoPep-1 probe (Xiangguo Che, Lianhua Chi, Clara Yongjoo Park, Gyoung-Ho Cho, Narae Park,
Seong-Gon Kim, Byung-Heon Lee, Je-Yong Choi, Arthritis Research & Therapy, 2015, 17:309)
- In Vivo Near-Infrared Fluorescence Imaging of Apoptosis Using Histone H1-Targeting Peptide Probe after
Anti-Cancer Treatment with Cisplatin and Cetuximab for Early Decision on Tumor Response (Hyun-
Kyung Jung, Kai Wang, Min Kyu Jung, In-San Kim, Byung-Heon Lee, PLoS ONE, 2014, 9(6): e100341)
- In vivo imaging of myocardial cell death using a peptide probe and assessment of long-term heart function
(Bodhraj Acharya, Kai Wang, In-San Kim, WoongChol Kang, Chanil Moon, Byung-Heon Lee, Journal of
Controlled Release, 2013, Volume 172, Issue 1, Pages 367–373)
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NpFlamma® HGC series
Description
As people’s interest in cancer increase, the importance of imaging cancer in order to develop an
anticancer drug along with the diagnosis of cancer is increasing day by day. NpFlamma®HGC series is
the nano-particle complex of amphiphilic polymer and near-infrared phosphor that enable to
selectively accumulate in the cancer tissues due to high permeability of angiogenesis of cancer
tissues. Therefore, it is easy for passive targeting tumor imaging using near-infrared penetration. In
addition, it is possible to contrast blood vessel through injecting probe into the blood vessel.
NpFlamma®HGC Series inside super hydrophobic material enables to embed hydrophobic drug and
accordingly it can be used for efficient transfer of tumor drug through interlinking with passive
targeting action. NpFlamma® HGC Series is the nano-particle produced based on chitosan, which has
almost no toxicity and side effect, and also has a good merit having a long half-life, high stability and
high water solubility. It seems that the use of this product enables more effective optical image.
[Figure 14] An effective tumor imaging is available using NpFlamma® HGC products group.
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[Figure 15] NpFlamma® HGC products group can penetrate into tumor cell tissues to observe strong
fluorescence specifically to cancer cell
Product List
Cat. No. Product name Emission
color
ExMax
(nm) EmMax
(nm)
Common
filter set
Excitation
laser line Size
PNC1201 NpFlamma® HGC 648 ● Red 648 675 Cy®5 594, 633 nm 10 doses, 50 doses, 100 doses
PNC1401 NpFlamma® HGC 675 ● Far red 675 698 Cy®5.5 633, 680 nm 10 doses, 50 doses, 100 doses
PNC1301 NpFlamma® HGC 749 ● NIR 750 782 Cy®7 680 nm 10 doses, 50 doses, 100 doses
PNC1601 NpFlamma® HGC 774 ● NIR 777 802 Cy®7.5 785 nm 10 doses, 50 doses, 100 doses
PNC1801 NpFlamma® HGC ICG ● NIR 785 821 Cy®7.5 785 nm 10 doses, 50 doses, 100 doses
In-vivo Imaging Protocol
- The experiment procedure for in-vivo imaging of the product is proceeded as follows.
- Prepare the experiment through vortexing by putting DW or PBS into the NpFlamma®HGC
series powder (1does=120ug/100ul)
- As fluorescent substance is weak against the light, it must be stored in the condition
protected from light
- Mouse fur may cause scattering, absorption etc. of excitation light when optical imaging is in
process. Therefore, it is necessary to use nude mouse or remove the mouse fur for
preparation of the test.
- It is recommended to prepare 31G syringe as injection must be conducted through the vein
of mouse.
- Prepare 5 week-old mouse Balb/c-nude, male.
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- Inject SCC7 cell line (1x106/0.1ml) subcutaneous of Balb/c-nude mouse.
- When the volume of tumor becomes 60~80mm3, take a time-zero image of each subject
before injection of the agent.
- Inject NpFlamma®HGC series (120ug/100ul) intravenously into mouse.
- The optimal imaging time point is 1hr, 3hr, 6hr, 9hr and 24 hr post injection of the
NpFlamma®HGC series.
- When take a 24hr imaging post injection of NpFlamma®HGC series, extract major organs (e.g
liver, lung, spleen, kidney, heart, and tumor) and then take of ex-vivo imaging.
In-vivo Imaging Result (IVIS)
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[Figure 16] As result of conducting fluorescence observation for the mouse after injecting NpFlamma®
HGC products group up to max. 7 days, it was possible to confirm a strong fluorescence from cancer
tissues until the 7th day
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NpFlamma HGC 675 (n=5)
* 각 Organ 의 Liver 에 대한 형광 Ratio.
Liver Lung Spleen Kidney Heart Tumor
1 1 0.940 - 0.509 - 1.812
2 1 0.669 - - - 1.747
3 1 0.944 - - - 1.650
4 1 0.657 - - - 1.509
5 1 0.613 - - - 1.998
AngioSense 680EX (n=5)
* 각 Organ 의 Liver 에 대한 형광 Ratio.
Liver Lung Spleen Kidney Heart Tumor
1 1 0.325 - 0.406 0.226 1.634
2 1 0.431 - 0.368 0.237 1.329
3 1 0.358 - 0.324 0.203 1.022
4 1 0.276 0.158 0.330 0.214 1.058
5 1 0.305 0.168 0.373 0.209 1.072
NpFlamma HGC 749 (n=5)
* 각 Organ 의 Liver 에 대한 형광 Ratio.
Liver Lung Spleen Kidney Heart Tumor
1 1 1.632 - - - 2.271
2 1 0.879 - - - 2.292
3 1 0.834 - - - 2.171
4 1 0.682 - - - 2.762
5 1 1.476 - - - 1.955
AngioSense 750EX (n=5)
* 각 Organ 의 Liver 에 대한 형광 Ratio.
Liver Lung Spleen Kidney Heart Tumor
1 1 0.357 - 0.388 - 1.311
2 1 0.398 - 0.410 - 1.626
3 1 0.319 - 0.372 - 1.358
4 1 0.360 - 0.405 - 1.400
5 1 0.396 - 0.451 - 1.389
[Figure 17] As result of comparing ex-vivo image of organ after 24 hr for NpFlamma® HGC product and
the other company’s product, it was confirmed that tumor accumulating efficiency against the other’s
organ was high against the other ’s tested organ, and that the accumulating trend of the other company’s
tested organ was low
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References
- New generation of multifunctional nanoparticles for cancer imaging and therapy (Kyeongsoon Park,
Seulki Lee, Eunah Kang, Kwangmeyung Kim, Kuiwon Choi, Ick Chan Kwon, Advanced Functional
Materials, 2009, Volume 19, Issue 10, Pages 1553–1566)
- Tumor-homing multifunctional nanoparticles for cancer theragnosis: Simultaneous diagnosis, drug delivery,
and therapeutic monitoring (Kwangmeyung Kim, Jong Ho Kim, Hyungkyu Park, Yoo-Shin Kim,
Kyeongsoon Park, Heayun Nam, Seulki Lee, Jae Hyung Park, Rang-Woon Park, In-San Kim, Kuiwon
Choi, Sang Yoon Kim, Kinam Park, Ick Chan Kwon, Journal of Controlled Release, 2010, Volume 146,
Issue 2, Pages 219–227)
- Glycol chitosan nanoparticles as specialized cancer therapeutic vehicles: Sequential delivery of
doxorubicin and Bcl-2 siRNA (Hong Yeol Yoon, Sejin Son, So Jin Lee, Dong Gil You, Ji Young Yhee, Jae
Hyung Park, Maggie Swierczewska, Seulki Lee, Ick Chan Kwon, Sun Hwa Kim, Kwangmeyung Kim &
Martin G. Pomper, Scientific Reports, 2014, 4, Article number: 6878)
NpFlamma® MMP series
Description
Cancer cell is influenced by ECM (extracellular matrix), ECM-related growth factors and cytokine
and surrounding cells in the growth process of cancer cell. Especially four characteristics of cancer
such as migration, invasion, metastasis and angiogenesis react according to the surrounding micro
environment. In this case, MMP (matrix metalloprotease) that is the protease controls cell-cell and
cell-ECM interactions playing an important role in the growth of cancer cell. Accordingly a number of
researches have been proceeding a variety of researches using MMP. As this product generates
fluorescence according to the absolute volume of MMP, it can be effectively accumulated in the
cancer tissues and it is possible for optical imaging of the tissues where MMP is over-expressed. As
this product enables to quickly check activity and suppression of protease through imaging, which
can be applied to drug screening, and it is possible for real-time based cell imaging and non-invasive
tissues imaging for cell and tissue. Especially in addition to the method of screening new drug such
as inhibitor that inhibits over-expression of protease, it can be used usefully for early diagnosis of a
variety of diseases and incurable diseases like autoimmune diseases that are MMP-related cancer,
ostarthritis, rheumarthritis, dementia, etc. In addition, this products group is expected to be easy to
use owing to have various fluorescent wavelengths.
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[Figure 18] Operation principle of NpFlamma® MMP Series and high reaction to Enzyme were
confirmed
[Figure 19] High restoring force of fluorescence of NpFlamma® MMP Series according to conc. of
Trypsin was confirmed
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Product List
Cat. No.
Product name Emission
Color
ExMax
(nm)
EmMax
(nm)
Common
filter set
Excitation
laser line
Size
PNM0101 NpFlamma® MMP-2,9 ICG NIR 785 812 Cy®7.5 785 nm 10 dose, 50 dose, 100 dose
PNM0103 NpFlamma® MMP-2,9 648 Red 648 663 Cy®5 594, 633 nm 10 dose, 50 dose, 100 dose
PNM0104 NpFlamma® MMP-2,9 675 Far red 675 691 Cy®5.5 680 nm 10 dose, 50 dose, 100 dose
PNM0105 NpFlamma® MMP-2,9 749 NIR 749 774 Cy®7 785 nm 10 dose, 50 dose, 100 dose
PNM0106 NpFlamma® MMP-2,9 774 NIR 774 800 Cy®7.5 785 nm 10 dose, 50 dose, 100 dose
In-vivo Imaging Protocol
- The experiment procedure for in-vivo imaging of the product is proceeded as follows.
- Prepare the experiment through vortexing by putting DW or PBS into the NpFlamma® MMP
series powder (1does=120ug/100ul)
- As fluorescent substance is weak against the light, it must be stored in the condition
protected from light
- Mouse fur may cause scattering, absorption etc. of excitation light when optical imaging is in
process. Therefore, it is necessary to use nude mouse or remove the mouse fur for
preparation of the test.
- It is recommended to prepare 31G syringe as injection must be conducted through the vein
of mouse.
- Prepare 5 week-old mouse Balb/c-nude, male.
- Inject SCC7 cell line (1x106/0.1ml) subcutaneous of Balb/c-nude mouse.
- When the volume of tumor becomes 60~80mm3, take a time-zero image of each subject
before injection of the agent.
- Inject NpFlamma®MMP series (120ug/100ul) intravenously into mouse.
- The optimal imaging time point is 1hr, 3hr, 6hr, 9hr and 24 hr post injection of the
NpFlamma® MMP series
- When take a 24hours imaging post injection of NpFlamma® MMP series, extract major
organs (e.g liver, lung, spleen, Kidney, heart, and tumor) and then take of ex-vivo imaging.
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In-vivo Imaging Result (IVIS)
[Figure 20] As result of comparing the in-vivo images by hours for NpFlamma® MMP product and the
other’s product, high tumor accumulating efficiency against the other’s tested organ was confirmed
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[Figure 21] As result of comparing the ex-vivo images of organ between NpFlamma® MMP product and the
other’s product, high tumor accumulating efficiency against the other’s organ was confirmed
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References
- Optimization of matrix metalloproteinase fluorogenic probes for osteoarthritis imaging (Ju Hee Ryu,
Aeju Lee, Jin Hee Na, Seulki Lee, Hyung Jun Ahn, Jong Woong Park, Cheol-Hee Ahn, Byung-Soo Kim,
Ick Chan Kwon, Kuiwon Choi, Inchan Youn, Kwangmeyung Kim, Amino Acids, 2011, Volume 41, Issue 5,
pp 1113–1122)
- Dark Quenched Matrix Metalloproteinase Fluorogenic Probe for Imaging Osteoarthritis Development in
Vivo (Seulki Lee, Kyeongsoon Park, Seung-Young Lee, Ju Hee Ryu, Jong Woong Park, Hyung Jun Ahn,
Ick Chan Kwon, In-Chan Youn, Kwangmeyung Kim, Kuiwon Choi, Bioconjugate Chem., 2008, 19 (9), pp
1743–1747)
- Early Diagnosis of Arthritis in Mice With Collagen-Induced Arthritis, Using a Fluorogenic Matrix
Metalloproteinase 3–Specific Polymeric Probe (Ju Hee Ryu, Aeju Lee, Jun-Uk Chu, Heebeom Koo,
Chang-Yong Ko, Han Sung Kim, Soo-Young Yoon, Byung-Soo Kim, Kuiwon Choi, Ick Chan Kwon,
Kwangmeyung Kim, Inchan Youn, ARTHRITIS & RHEUMATISM, 2011, Vol. 63, No. 12, pp 3824–3832)
- Polymeric Nanoparticle-Based Activatable Near-Infrared Nanosensor for Protease Determination In Vivo
(Seulki Lee, Ju Hee Ryu, Kyeongsoon Park, Aeju Lee, Seung-Young Lee, In-Chan Youn, Cheol-Hee Ahn,
Soon Man Yoon, Seung-Jae Myung, Dae Hyuk Moon, Xiaoyuan Chen, Kuiwon Choi, Ick Chan Kwon,
Kwangmeyung Kim, Nano Lett., 2009, Vol. 9, No. 12, pp 4412-4416)
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Technical
assistance
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