Certificate of Analysis & Data Sheet

Mono-pegylated (with 20 kDa PEG) recombinant super active ovine 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 super active ovine leptin antagonist runs on the SGS-Page as 48 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. The half-life in circulation of pegylated recombinant super active ovine leptin antagonist after SC injection was over 20 hours. Recombinant super active ovine leptin antagonist was initially mutated, resulting in D23L/L39A/D40A/F41A mutant that was purified by proprietary chromatographic techniques. The pegylation of recombinant super active ovine leptin antagonist is similar to that described in Elinav et al. Endocrinology 150:3083-91 (2009) for pegylated recombinant super active mouse leptin antagonist

Escherichia coli

White lyophilized (freeze-dried) powder.

The pegylated recombinant super active ovine leptin antagonist was lyophilized from a concentrated (0.65mg/ml) solution with 0.003mM NaHCO3.

It is recommended to reconstitute the lyophilized pegylated recombinant super active ovine 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.

Lyophilized pegylated recombinant super active ovine leptin antagonist although stable at room temperature for several weeks, should be stored desiccated below - 20C. Upon reconstitution of pegylated recombinant super active ovine leptin antagonist at > 0.1 mg/ml and up to 2 mg/ml and filter sterilization pegylated recombinant super active ovine leptin antagonist can be stored at 4C or even room temperature for several weeks making it suitable for long term infusion studies using osmotic pumps. At lower concentration of pegylated recombinant super active ovine leptin antagonist addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.

The purity od pegylated recombinant super active ovine leptin antagonist is greater than 95.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.

The sequence of the N-terminal amino acids of the pegylated recombinant super active ovine leptin antagonist could not be determined indicating that the PEG moiety is attached to the N-terminus.

The purified pegylated recombinant super active ovine leptin antagonist consists of > 90% monomers as determined by gel-filtration chromatography.

PLR’s pegylated recombinant super active ovine 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 recombinant super active ovine leptin antagonist is 6-8 fold lower than the non-pegylated antagonist but is 15 fold higher as compared to pegylated recombinant super active ovine leptin antagonist

The endotoxin content of pegylated recombinant super active ovine leptin antagonist is less than 0.01 ng/µg (0.1 EU/µg of protein) .

Protein quantitation of the pegylated recombinant super active ovine leptin antagonist was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.200 as the extinction coefficient for a 0.1% (1mg/ml) protein solution at pH 8.0. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

PLR's pegylated recombinant super active ovine leptin antagonist is furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives, local application or household chemicals. It is for Research Purposes Only – Commercial Use has been Licensed Exclusively to a Third Party.