Journal
JOURNAL OF BIOACTIVE AND COMPATIBLE POLYMERS
Volume 25, Issue 1, Pages 75-97Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1177/0883911509354230
Keywords
PEGylation; melittin; hemolysis; blood coagulation; bioconjugation
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To promote and understand the structure-property relationship for hemostasis, we modified melittin (MLT) using a four-arm poly(ethylene glycol) (PEG) with N-hydroxysuccinimide ester. The PEGylation was characterized by FTIR, MALDI-MS, NMR, a bicinchoninic acid assay, circular dichroism, hemolysis assay, and thromboelastography. Changes in the reaction conditions affected the extent of the modification, the numbers of MLT conjugated to PEG arms, and possible PEGylation sites. The reaction at pH 9.2 with a high MLT/PEG ratio, resulted in the highest modification. Reactions in dimethylsulfoxide (DMSO) resulted in more multi-arm coupled MLT, reaching a maximum of four MLT per PEG. The helicity of the modified peptide, relative to the native peptide, was essentially maintained in DMSO, but substantially lost at pH 9.2. PEGylation reduced the hemolytic effects of MLT and subsequently changed its coagulation profiles. The overall hemostatic effects of MLT modified in DMSO indicate that this may be a convenient approach to the PEGylation of biomolecules for biomedical applications.
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