Leptine, gepegyleerd

Mono-pegylated (with 20 kDa PEG) recombinant rat leptin is an one polypeptide chain containing 146 amino, an additional Ala at N-terminus and one molecule of PEG 20 kDa at its N-terminus, having an expected molecular mass of ~ 35.6 kDa as determined by MS. However due to enlarged hydrodymanic volume it runs on the SDS-PAGE as 48 kDa protein and in gel-filtration on Superdex 200 as over 100 kDa protein. The half-life in circulation of pegylated recombinant rat leptin after SC injection was over 20 hours. Recombinant rat leptin as purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136 and then pegylated.

Lep; ob; obese

25608

P50596

NP_037208

NM_013076

Rat

Rat

PEG

AVPIHKVQDDTKTLIKTIVTRINDISHTQSVSARQRVTGLDFIPGLHPILSLSKMDQTLAVYQQILTSLPSQNVLQIAHDLENLRDLLHLLAFSKSCSLPQTRGLQKPESLDGVLEASLYSTEVVALSRLQGSLQDILQQLDLSPEC

It is recommended to reconstitute the lyophilized recombinant rat leptin in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml, which can then be further diluted with other aqueous solutions.

< 0.1 ng/µg of protein (< 1EU/µg)

> 98.0% as determined by Gel filtration and SDS-PAGE gel.

Pegylated recombinant rat leptin is capable of stimulating proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. Its in vitro activity is only slightly lower than the non-pegylated recombinant rat leptin but in vivo it has profound weight reducing effect (as compared to the non-pegylated leptin), resulting mainly from reduced food intake.

146

Lyophilized

Water

35.6 kDa

E. coli