Leptine gepegyleerde superantagonist (mutant D23L/L39A/D40A/F41A)

Mono-pegylated mouse super leptin antagonist (with 20 kDa PEG) 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 super mouse leptin antagonist runs on the SDS-PAGE as a 55 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. Pegylated recombinant super mouse leptin antagonist half-life in circulation after SC injection was over 20 hours. Super mouse leptin antagonist (SMLA) was initially mutated, resulting in D23L/L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques. Its pegylation is similar to that described in Elinav et al. Endocrinology 150:3083-91 (2009) for pegylated recombinant mouse leptin antagonist.

Lep; ob; obese

16846

P41160

NP_032519.1

NM_008493.3

Mouse

Mouse

PEG

AVPIQKVQDDTKTLIKTIVTRINLISHTQSVSAKQRVTGAAAIPGLHPILSLSKMDQTLAVYQQVLTSLPSQNVLQIANDLENLRDLLHLLAFSKSCSLPQTSGLQKPESLDGVLEASLYSTEVVALSRLQGSLQDILQQLDVSPEC

It is recommended to reconstitute the lyophilized pegylated recombinant super mouse 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)

> 95.0% as determined by Gel filtration and SDS-PAGE gel.

Pegylated recombinant super mouse leptin antagonist is capable of inhibiting leptin-induced proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. Pegylated recombinant super mouse leptin antagonist in vitro activity is 6-8 fold lower than the non-pegylated pegylated recombinant super mouse leptin antagonist but in vivo it has profound weight gain effect (as compared to the non-pegylated antagonist), resulting mainly from increased food intake. Its in vivo activity compared to that of PEG-MLA is 9-27 fold higher.

146

Lyophilized

Water

35.6 KkDa

E. coli

AVPIQ