Certificate of Analysis & Data Sheet

Chicken recombinant growth hormone (chGH) mutein G119R produced in E.Coli (22 K) is a single, non-glycosylated, polypeptide chain containing 191 amino acids with an additional Ala at its N-terminus and having a molecular mass of 22.3 kDa. chicken recombinant growth hormone G119R is purified by proprietary chromatographic techniques. For reference see Eliasiewicz et al. (2006) Ann N Y Acad Sci. 1091:501-508.

E. coli

White lyophilized (freeze-dried) powder.

The chicken recombinant growth hormone was lyophilized from a concentrated (1mg/ml) solution with 0.03% NaHCO3 adjusted to pH 8.

It is recommended to reconstitute the lyophilized chicken recombinant growth hormone mutant in sterile water or 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml and not more than 3 mg/ml, which can then be further diluted to other aqueous solutions, preferably in presence of carrier protein.

Lyophilized chicken recombinant growth hormone G119R although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution of chicken recombinant growth hormone mutant at > 0.1 mg/ml andup to 3 mg/ml and filter sterilization chicken recombinant growth hormone mutant can be stored at 4C for several weeks. At lower concentration addition of a carrier protein (0.1% HSA or BSA) is recomended. Please prevent freeze-thaw cycles.

The purity of chicken recombinant growth hormone mutant is greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
(c) Analysis by RP-HPLC.

The sequence of the first five N-terminal amino acids of chicken recombinant growth hormone mutant was determined and was found to be Ala-Thr-Phe-Pro-Ala.

The purified chicken recombinant growth hormone G119R (22K) consists of > 99% monomers as determined by gel-filtration chromatography.

Chicken recombinant growth hormone G119R mutant did not bind to ovine growth hormone extracellular domain (ECD) and was devoid of any biological activity in FDC-P1 3B9 cells. However, in binding experiments that were carried out using chicken liver membranes, both ovine growth hormone and chicken recombinant growth hormone showed similar IC50 values in competition with 125I-oGH, while the IC50 of chicken recombinant chicken growth hormone G119R mutein was 10-fold higher. These results emphasize the importance of species specificity and indicate the possibility of antagonistic activity of chicken recombinant growth hormone G119R in homologous system.

Less than 0.1 ng/µg (IEU/µg) of chicken recombinant growth hormone G119R (22 K).

Protein quantitation of chicken recombinant growth hormone mutant was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.74 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0 This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

The chicken recombinant growth hormone mutant is offered by PLR for research, laboratory or further manufacturing purposes.