Leptinegepegyleerde antagonist (mutant L39A/D40A/F41A)

Mono-pegylated (with 20 kDa PEG) recombinant rat leptin antagonist is 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 pegylated recombinant rat leptin antagonist runs on the SDS-PAGE as 48 kDa protein and on gel-filtration Superdex 200 column as over 100 kDa protein. The half-life of recombinant rat leptin antagonist in circulation after SC injection was over 20 hours. Recombinant rat leptin was initially mutated, resulting in L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136.

Lep; ob; obese

25608

P50596

NP_037208

NM_013076

Rat

Rat

PEG

AVPIHKVQDDTKTLIKTIVTRINDISHTQSVSARQRVTGAAAIPGLHPILSLSKMDQTLAVYQQILTSLPSQNVLQIAHDLENLRDLLHLLAFSKSCSLPQTRGLQKPESLDGVLEASLYSTEVVALSRLQGSLQDILQQLDLSPEC

It is recommended to reconstitute the pegylated rat leptin antagonist 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)

> 99.0% as determined by Gel filtration and SDS-PAGE gel.

Pegylated rat leptin antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. The in vitro activity of pegylated rat leptin antagonist is 5-6 fold lower than the non-pegylated antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated antagonist), resulting mainly from increased food intake.

146

Lyophilized

Water

35.6 kDa

E. coli

AVPIQ