NOF produces the world’s best quality Activated PEGs from our high-quality methoxypoly(ethylene glycol) (mPEG). NOF also supplies our proprietary high-purity branched, multi-arm and hetero-functional Activated PEGs tailored to meet our customers’ PEG Drugs. NOF can supply Activated PEGs produced under GMP standards in our state-of-the-art GMP facility. NOF can help you to modify your drugs by using our PEGylation technology and our high-quality Activated PEGs.
NOF has been producing a high-quality activated PEGs, SUNBRIGHT series, which possess the most suitable quality for preparation of physiologically active drug and biologics. SUNBRIGHT series are characterized as extremely high-purity PEGs with a variety of functional groups, whereby the products of this series are recognized as optimal PEGylation drugs. As illustrated in this figure, application of the activated PEGs with various functional groups enable introduction of the PEG chains into drugs, enzymes, phospholipids and other biologics. Covalent conjugation of hydrophobic macromolecules with activated PEGs leads to the formation of macromolecular micelles (Polymer Micelles), which allow homogeneous dispersion of hydrophobic drugs in aqueous media. Furthermore, when the PEGylated phospholipids are used as liposomes, the aqueous layer can be added to its surface, with homogeneous dispersal of the liposome preparations encapsulating drugs within them.
As stated above, PEGylation plays an important role in the stabilization of drugs, reduction of their antigenicity and decrease in the drug doses, besides augmenting the targeting ability via binding biologics onto their surfaces. We can do supply our activated PEGs from small scale reagent grade to huge amount GMP grade with high purity.
PEGylation technology is applied for biologics modifications such as cytokines, antibody fragment, interleukins, hormones, oligonucleotides and some of proteins and peptides.
PEGylation can be effected to
Prolonging Blood Circulation
NOF provides high-performance activated PEGs for PEGylated drugs from early development stage to commercial use with our 20 years of experience of manufacturing high-quality methoxy polyethylene glycol, which is a starting material of activated PEG for pharmaceutical area. Our activated PEGs have a narrow polydisperity and low diol content with wide range of molecular weight, from 2kDa to 80kDa.
Analysis of PEG-ALD-Insulin by SDS-PAGE with Coomassie brilliant blue. Insulin was reacted with NOF PEG-ALD at 4°C. The PEGylated insulin was purified on Q Sepharose Fast Flowcolumn.
lane M molecular weight markers
lane 1 Insulin
lane 2 PEG(ME-050AL)-insulin eluate
lane 3 PEG(ME-100AL)-insulin eluate
lane 4 PEG(ME-200AL)-insulin eluate
lane 5 PEG(ME-300AL)-insulin eluate
Coupling of PEG derivatives to Proteins: examples of conditions
Coupling of PEG-NHS derivatives to protein amines (PEG-NHS + Protein-NH 2) Conditions 1: 50 mM phosphate buffer (pH 7.2), 4℃, 6 hrs *1)
Conditions 2: Borate-phosphate buffer (pH 8.0), 25℃, 2 hrs *2)
Coupling of PEG-Aldehyde derivatives to the NH 2 group of proteins (PEG-Ald + Protein-NH 2)
Conditions : sodium cyanoborohydride (10 eq . .), 4 deg C, 20 hrs *3)
Coupling of PEG-Maleimide derivatives to the SH group of proteins (PEG-Maleimide + Protein-SH)
Conditions: 100 mM phosphate buffer (pH 6.5), 4 deg C, 4 hrs
Coupling of PEG-NH 2 derivatives to the COOH group of proteins (PEG-NH 2 + Protein-COOH)
Conditions: 50mM phosphate buffer (pH 7.2), WSC (2 eq.), 4 deg C, 10 hrs
Coupling of PEG-p-Nitrophenyloxycarbonyl derivatives to the NH 2 group of proteins (PEG-NP + Protein-NH 2)
Conditions: borate-phosphate buffer (pH 8.0-8.3), r.t, overnight *2)
*1) Abuchowski, A., et al. (1984) Cancer Biochem. Biophys. 7, 175.
*2) Sartore, L., et al. (1991) Appl. Biochem. Biotechnol. 27, 45
*3) US 5,824,784
We supply different series of PEG derivatives with various versatile functional groups.
Purity of NOF’s activated products
We use our advanced technologies to measure the quality of our activated PEGs. The following are the typical analysis data for each type of activated PEGs which are measured by NMR or HPLC methods. Please note that those numbers are not specification of the product.
CS type mPEG-succinimidyl succinate (Mw 2k and 5k) : activity over 95% (NMR)
GS type mPEG-succinimidyl glutarate (Mw 5k, 10k, 20k, 30k and 40k) : activity around 95% (NMR)
AS type mPEG-succinimidyl methyl ester (Mw 2k and 5k) : activity around 95% (NMR)
HS type mPEG-succinimidyl pentyl ester(Mw 5k, 10k, 20k, 30k and 40k): activity around 95% (NMR)
TS type mPEG-succinimidyl carbonate(Mw 5k, 10k, 12k, 20k, 30k and 40k): activity over 95% (HPLC)
pNP type (NP series)
mPEG-p-nitrophenyl (Mw 2k, 5k, 10k, 20k, 30k and 40k): activity over 95% (HPLC)
Aldehyde type(AL series)
mPEG-propylaldehyde (Mw 5k, 10k, 20k, 30k and 40k): activity over 85% (HPLC). The activity for Mw20k, it is over 92% as actual data by HPLC.
PA type mPEG-propylamine (Mw 2k, 5k, 10k, 12k, 20k, 30k and 40k): activity over 95% (HPLC)
EA type mPEG-ethylamine (Mw 5k, 10k, 20k, 30k and 40k): activity over 95% (HPLC)
thiol type(SH series)
mPEG-thiol (Mw 2k, 5k, 10k, 20k, 30k and 40k): activity around 95% (HPLC)
Maleimide type(MA series)
mPEG-maleimide (Mw 2k, 5k, 10k, 20k, 30k and 40k): activity around 95% (HPLC)
Oxyamine type(CA series)
mPEG-oxyamine (Mw 10k, 20k, 30k and 40k): activity around 95% (HPLC)
Iodoacetoamide type(IA series)
mPEG-irdoacetoamide (Mw 20k, 30k and 40k): activity around 90% (HPLC)
NOF has more than 20 years history for manufacturing of high-quality mPEGs and Activated PEGs. These experiences enable NOF to have thorough knowledge of PEGs and considerable achievement in analysis of Activated PEGs. We also have invented innovative analytical methods for assaying purity of activated PEGs and for determining impurity levels. (*Patent No.: JP4543917) Previously, the activity for activated PEGs is measured by NMR. However, this method is not very good detection due to polymer, especially for high molecular weight. We developed the analysis method by using HPLC. As the result of the analysis, the activity sometime seems lower than the result of NMR because main peak can completely separate from non-active peak and activated impurity peaks (like dimer).
HPLC analysis for M-PEG-COOH
GPC Analysis of M-PEG5K and M-PEG-30K
MALDI-TOF-MS Analysis of M-PEG5K and M-PEG-30K
cGMP Manufacturing Facilities
NOF Activated PEGs, SUNBRIGHT® series are produced in facilities with state-of-the-art technology, operated under cGMP. The cGMP facilities are capable of producing small to large batch sizes, using proprietary know-how on scale-up procedure, depending on customers’ needs. These cGMP facilities have been audited by pharmaceutical companies throughout the world. NOF receives high reputation from our customers.
Research & Development
Our R&D facility was opened in December 2005. This allows us to continue our development of novel activated PEGs and technologies for PEGylation.
Research & DevelopmentOur R&D facility was opened in December 2005. This allows us to continue our development of novel activated PEGs and technologies for PEGylation.
NOF has over 20 years of experience in supplying high-quality methoxy polyethylene glycol (mPEG) for pharmaceutical market. The mPEG is starting material of PEG derivatives, which are patented through our proprietary technology. Based on extensive range of technological know-how and capabilities, NOF offers custom manufacturing of Activated PEGs from early stage development to commercial scale. Sizewise, we can offer gram size synthesis to kg size for non GMP product. We also offer cGMP custom synsthesis.
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1. Mono-functional Linear PEGs
2. Bi-Functional PEGs
3. Multi-Arm PEGs
4. Branched PEGs
5. Forked PEGs
6. Heterofunctional PEGs
7. Single Molecule PEGs PUREBRIGHT® Series
8. Poly-Hydroxyl Activated PEGS
10. Biodegradable Polymers for gene / nucleic acid Delivery SUNBRIGHT® ME-DT & DT series
11. Biodegradable block polymer for drug delivery
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Molecular weight: 1555.66
Molecular weight: 1530.65
Molecular weight: 1717.85
Molecular weight: 1581.65
X = 8 Molecular weight: 441.51
X=12 Molecular weight: 617.72
X=8 Molecular weight: 541.63
X=8 Molecular weight: 717.84
X=8 Molecular weight: 592.63
X=8 Molecular weight: 689.71
X=12 Molecular weight: 768.84
X=12 Molecular weight: 865.92