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Custom Vector Construction> | UNIPROT ID #O60894) was synthesized in vitro and cloned into the bacte- rial protein expression vector pET28b (Novagen/EMD Mil- lipore) between the NdeI and EcoRI sites by the GenScript Corporation (Piscataway | Get A Quote |
We modified amylin chemically by conjugating methoxyl polyethyleneglycol succinimidyl carbonate (mPEGSC) of varying molecular weights (1 kDa, 2 kDa and 5 kDa). The reaction occurred within a few minutes, resulting in at least four distinct PEGylated products. The reaction products were separated by reversed-phase chromatography and identified by mass-spectrometry. The monoPEGylated and diPEGylated amylin products were generated rapidly through conjugation to the two amino groups of the N-terminal lysine residue. Both PEGylated amylin products bound to the receptor activity-modifying protein 1 (RAMP1). Pharmacological evaluation by subcutaneous administration in mice of monoPEGylated and diPEGylated amylin o... More