Volume 8, Issue 31 (7-2018)
NCMBJ 2018, 8(31): 95-103 |
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Akrami M, Noofeli M, Tabatabaei M S. OPTIMIZATION OF DIPHTHERIA TOXOID PRECIPITATION BY USING SATURATED AMMONIUM SULPHATE IN ORDER TO IMPROVE DIPHTHERIA VACCINE . NCMBJ 2018; 8 (31) :95-103
URL: http://ncmbjpiau.ir/article-1-1133-en.html
URL: http://ncmbjpiau.ir/article-1-1133-en.html
Department of Biology,Faculty of Science,Islamic Azad University Central Tehran Branch,Iran
Abstract: (5178 Views)
Aim and Background: Diphtheria toxin is an extracellular protein of Corynebacterium diphtheria that inhibits protein synthesis and kills susceptible cells. Ammonium sulfate is an inorganic salt that is used to purify and precipitate proteins, which, by increasing salt content, decreases the solubility of proteins, and proteins interconnect and precipitate through their hydrophobic parts. Diphtheria vaccine is prepared by purifying and treating toxoid diphtheria bacteria, which is one of the methods of treating, using ammonium sulfate as a dry salt or preparing an over-saturated solution.
Material and Methods: Diphtheria bacteria grown in optimal conditions and increased nutrient and toxin production by bacteria and inactivating the toxin into toxoid, in the manufacture of the vaccine is use. All media must be at least 24 hours at a temperature of 37-35°C will be incubated. In order to bacterial toxoid deposition of supersaturated solution 4.32 M ammonium sulfate with different percentages were used. The results of this procedure were investigated using SDS PAGE and Lf TEST ( Lateral Flow Test).
Results: Results revealed the use of 4.32 M supersaturated ammonium sulfate instead of the use of dry salt of ammonium sulfate, used in the traditional method (WHO TRS, No. 980), used previously in the vaccine industry, Eliminating the long-term dialysis phase On the other hand, the final product (toxoid) has been tediously reduced It also removes unwanted impurities from the finished product And has made the net product even more scalable.
Conclusion: Since the purpose of vaccine production is industry to achieve maximum toxoid of the bacteria, so use the procedure in this research project (optimizing of DT toxin precipitation by 4.32 M super saturation of ammonium sulphate) , in addition to reducing the loss of the final product and remove the prolonged dialysis stage and so the economy will become more affordable.
Material and Methods: Diphtheria bacteria grown in optimal conditions and increased nutrient and toxin production by bacteria and inactivating the toxin into toxoid, in the manufacture of the vaccine is use. All media must be at least 24 hours at a temperature of 37-35°C will be incubated. In order to bacterial toxoid deposition of supersaturated solution 4.32 M ammonium sulfate with different percentages were used. The results of this procedure were investigated using SDS PAGE and Lf TEST ( Lateral Flow Test).
Results: Results revealed the use of 4.32 M supersaturated ammonium sulfate instead of the use of dry salt of ammonium sulfate, used in the traditional method (WHO TRS, No. 980), used previously in the vaccine industry, Eliminating the long-term dialysis phase On the other hand, the final product (toxoid) has been tediously reduced It also removes unwanted impurities from the finished product And has made the net product even more scalable.
Conclusion: Since the purpose of vaccine production is industry to achieve maximum toxoid of the bacteria, so use the procedure in this research project (optimizing of DT toxin precipitation by 4.32 M super saturation of ammonium sulphate) , in addition to reducing the loss of the final product and remove the prolonged dialysis stage and so the economy will become more affordable.
Type of Study: Research Article |
Subject:
Cellular and molecular
Received: 2018/09/8 | Accepted: 2018/09/8 | Published: 2018/09/8
Received: 2018/09/8 | Accepted: 2018/09/8 | Published: 2018/09/8
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