Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Hormones – e.g. – prolactin – thymosin – growth factors – etc.
Reexamination Certificate
1996-06-03
2001-10-23
Sayala, Chhaya D. (Department: 1761)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Hormones, e.g., prolactin, thymosin, growth factors, etc.
C530S416000, C530S417000
Reexamination Certificate
active
06307031
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention relates to the discovery of non-toxic beta taipoxin as a cell growth factor and a potent mitogen from poisonous snake venom. Said composition consists of a polypeptide of beta taipoxin having molecular weight approximately 14,000 daltons and is free of toxic effects.
Taipoxin as a whole intact molecule isolated from the venom of the Australian taipan Oxyuranus s. scutellatus is the most lethal neurotoxin. The whole molecule of taipoxin is a complex, composed of three subunits designated as alpha, beta and gamma, having molecular weight 45,6000 daltons. Taipoxin can not be isolated by ion-exchange chromatography, since ion exchangers tend to dissociate the active toxin complex. The major lethality of taipoxin is due to the very basic alpha subunit. The molecular weights of alpha, beta and gamma subunits of taipoxin are 13,750, 13,473 and 18,354 daltons respectively.
To date numerous growth factors have been isolated from various sources, and have been characterized. To name a few, epidermal growth factor (EGF), fibroblast growth factor (FGF), nerve growth factor (NGF) and platelet derived growth factor (PDGF).
It is an object of this invention that the beta taipoxin as a cell growth factor can be used to grow cells in serum free medium. Routinely, cells are grown in the presence of 5 to 20% fetal bovine serum (FBS) for research or production. The purification of products derived from cells grown in serum containing medium is a cumbersome task and furthermore, fetal bovine serum is the most expensive ingredient of the medium. A mitogen like cell growth factor will provide serum free environment with adequate cell proliferation.
A further object of this invention is to provide non-toxic beta taipoxin having mitogenic activity as a composition for the promotion of rapid burn and wound healing. In the case of cuts, surgical incisions, and abrasions to lessen the risk of infection and to shorten recovery time.
SUMMARY OF THE INVENTION
Cell growth factor has been isolated as the beta subunit of taipoxin by fractionating Oxyuranus s. scutellatus snake venom by high pressure liquid chromatography. After testing for mitogenic activity in all fractions, the mitogenic activity was revealed in one of the fractions. Cell growth factor is a beta taipoxin, which is a non-toxic fraction of snake venom and is a potent mitogen. Cell growth factor has been isolated from the venom of the poisonous snake species Oxyuranus s. scutellatus.
The concentration of 0.1 &mgr;g/ml of the beta taipoxin non-toxic subunit becomes a cell growth factor and in serum free medium it gives growth equivalent to medium containing 10% serum for a wide range of cells. Cell growth factor is a beta taipoxin peptide having a molecular weight 14,000 daltons.
In vivo experiments proved that beta taipoxin as a cell growth factor acts as a potent mitogen. The cut portion of mouse hip skin healed much faster when cell growth factor was applied as compared to the control counter part. Cell growth factor helped heal a chronic foot wound of a friend. Cell growth factor is claimed as wound healer and its use may be extended to treat burns.
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Casperson John R.
Sayala Chhaya D.
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