Leptinegepegyleerde antagonist (mutant L39A/D40A/F41A)

Mono-pegylated (with 20 kDa PEG) recombinant mouse 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 mouse pegylated leptin antagonist runs on the SDS-PAGE as 48 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. Mouse pegylated leptin antagonist half-life in circulation after SC injection was over 20 hours.. Mouse leptin antagonist 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 and its pegylation is described in Elinav et al. Endocrinology 150:3083-91 (2009) .

Lep; ob; obese

16846

P41160

NP_032519.1

NM_008493.3

Mouse

Mouse

PEG

AVPIQKVQDDTKTLIKTIVTRINDISHTQSVSAKQRVTGAAAIPGLHPILSLSKMDQTLAVYQQVLTSLPSQNVLQIANDLENLRDLLHLLAFSKSCSLPQTSGLQKPESLDGVLEASLYSTEVVALSRLQGSLQDILQQLDVSPEC

It is recommended to reconstitute the lyophilized recombinant mouse pegylated 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)

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

Recombinant mouse pegylated 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 the pegylated recombinant mouse leptin antagonistis 6-8 fold lower than the non-pegylated antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated recombinant mouse leptin antagonist), resulting mainly from increased food intake.

146

Lyophilized

Water

35.6 KkDa

E. coli

AVPIQ