Certificate of Analysis & Data Sheet

Recombinant common carp (Cyprinus carpio) growth hormone (caGH) produced in E.Coli (21.4 K) is a single, non-glycosylated, polypeptide chain containing 188 amino acids and having a molecular mass of 21404 Da. Recombinant common carp growth hormone is purified by chromatographic techniques, according to Fine et al. General and Comparative Endocrinology, 89,51-61 (1993).

E. coli

White lyophilized (freeze-dried) powder.

The recombinant common carp 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 recombinant common carp growth hormone in sterile water or 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml and not more than 3 mg/ml. Recombinant common carp growth hormone can then be further diluted to other aqueous solutions, preferably in presence of carrier protein.

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

The purity od recombinant common carp growth hormone is greater than 95.0% 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 seven N-terminal amino acids of recombinant common carp growth hormone was determined and was found to be Ser-Asp-Asn-Gln-Arg-Leu-Phe.

The purified recombinant common carp growth hormone (21.4K) consists of > 98% monomers as determined by gel-filtration chromatography.

Recombinant common carp growth hormone is biologically active in rat 3T3 F442A preadipocytes, though its activity is 15-fold lower compared to bovine GH, but it is equally potent in vivo in promoting carp growth (Fine et al.1993). Recombinant common carp growth hormone is capable to form a 1:2 complex with oGHR-ECD and induce proliferation of PFD-P1 cells stably transfected with rabbit growth hormone receptor, though its activity is 5-fold lower compared to bovine GH (A. Gertler, unpublished data).

Less than 0.1 ng/µg (IEU/µg) of recombinant common carp growth hormone.

Protein quantitation of recombinant common carp growth hormone was carried out by two independent methods:
UV spectroscopy at 280 nm using the absorbency value of 0.47 as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0 This value is calculated by DNAman computer analysis program of protein sequences.

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