Tetanus toxoid purification: a case study.
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Filtration↗
Biomedical subjects
F Fratelli
Publications and source records attributed to F Fratelli.
Specific peptides of casein pancreatic digestion enhance the production of tetanus toxin.
Casein pancreatic digest is the basic bacterial growth medium used for diphtheria, botulinum and tetanus toxin vaccine production. It is known that the variation in the peptide content of the casein digest directly affects final toxin yields. In this study, the identification and sequences of eight peptides, four to eight amino acids in length, of casein pancreatic digestion, which seem to be involved in the enhancement of tetanus toxin production, are described. They all contain one or two residues of proline/molecule and a predominance of hydrophobic amino acid residues. The most active peptides show a general structure of Pro-aromatic-Pro, and this pattern resembled the motif displayed by bradykinin-potentiating peptides found in snake venoms. By analogy with the mechanism of bradykinin potentiation through inhibition of the proteolytic degradation of bradykinin, it is suggested that the six peptides identified here could protect the tetanus toxin from proteolysis, once secreted by the bacteria.
Animals↗
Development and validation study for the chromatographic purification process for tetanus anatoxin on Sephacryl S-200 High Resolution.
The tetanus purified anatoxin is used in the preparation of multiple immunoprophylactics. WHO (World Health Organization) specifies that the tetanus anatoxin must exhibit a degree of purity greater than or equal to 1,000 Lf/mg protein nitrogen (PN). Today liquid chromatography is a well established technique for the purification of tetanus anatoxin and several different methods are used in production scale. On a small scale, we purified tetanus anatoxin on Sephacryl S-200 High Resolution (gel filtration) and we obtained a successful high-yield purification. On the basis of these results, by combining conventional tangential flow filtration (TFF) at 50,000 N.M.W.L. (Nominal Molecular Weight Limit) ultrafiltration membrane with gel filtration on Sephacryl S-200 High Resolution, we have been able to purify 14 lots of tetanus anatoxin using the Bioprocess System (Amersham Pharmacia Biotech) to a large scale operation. Using this method, 77,401,332 doses of tetanus toxoid were prepared in 14 consecutive lots, supporting the reproducibility and reliability of the method presented here.
Acrylic Resins↗