A new method to increase selectivity of transglutaminase mediated PEGylation of salmon calcitonin and human growth hormone
| Autor: | Anna Mero, Mariano Schiavon, Francesco M. Veronese, Gianfranco Pasut |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Protein conjugation
Calcitonin Spectrometry Mass Electrospray Ionization Tissue transglutaminase Molecular Sequence Data Pharmaceutical Science Polyethylene glycol Polyethylene Glycols Substrate Specificity Rats Sprague-Dawley chemistry.chemical_compound PEG ratio TGase Animals Humans Technology Pharmaceutical Amino Acid Sequence chemistry.chemical_classification Drug Carriers Binding Sites Transglutaminases biology Chemistry Human Growth Hormone Circular Dichroism PEGylation Substrate (chemistry) Rats Enzyme Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Drug delivery biology.protein Solvents Calcium Female Conjugate |
| Zdroj: | Journal of controlled release : official journal of the Controlled Release Society. 154(1) |
| ISSN: | 1873-4995 |
| Popis: | Modification of therapeutic proteins and peptides by polyethylene glycol (PEG) conjugation is a well-known approach to improve the pharmacological properties of drugs. Several chemical procedures of PEG coupling are already in use but an alternative method based on microbial transglutaminase (mTGase) was recently devised. The enzyme catalyzes the link of mPEG-NH 2 to glutamines (Gln) of a substrate protein. In this case the advantage resides in the fact that usually only few Gln(s) in a protein are substrate of mTGase. In order to further restrict the selectivity of the enzyme, we investigated a new approach leading to the formation of a single conjugate isomer as well as for those proteins containing two or more Gln(s) as mTGase substrates. It was found that the addition of co-solvents in the reaction mixture influenced both the secondary structure of the targeted protein and the mTGase activity. The enzymatic PEGylation under these conditions yielded only mono- and selectively modified conjugates. The method was investigated with salmon calcitonin (sCT) and human growth hormone (hGH). In the case of sCT we also demonstrated the importance of site-selective conjugation for the preservation of in vivo activity. |
| Databáze: | OpenAIRE |
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