Leptine N82K-mutant, gepegyleerd

Mono-pegylated (with 20 kDa PEG) recombinant human leptin N82K mutant 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 recombinant human pegylated leptin runs on the SDS-PAGE as 48 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. Recombinant human pegylated leptin N82K mutant's half-life in circulation after SC injection was over 20 hours. Recombinant human leptin N82K mutant was purified by proprietary chromatographic techniques according to Salomon et al (2006) Protein Expression and Purification 47, 128–136 and then pegylated.

Lep; ob; obese

3952

P41159

NP_000221.1

NM_000230

Human

Human

PEG

AVPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVTGLDFIPGLHPILTLSKMDQTLAVYQQILTSMPSRNVIQISNDLEKLRDLLHVLAFSKSCHLPWASGLETLDSLGGVLEASGYSTEVVALSRLQGSLQDMLWQLDLSPGC

It is recommended to reconstitute the lyophilized recombinant human pegylated leptin N82K mutant 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.

Recombinant pegylated human leptin N82K mutant is less than 0.1% active active as evidenced by inducing proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor. This abolishment of activity results from drastically reduced affinity toward leptin receptor.

146

Lyophilized

Water

35.6 kDa

E. coli

AVPIQ