Golchinfar, F., Madani, R., Moazeni Jula, G. (2009). Purification & separation of Bacillus anthracis 34F2 Antigens with UF & HPLC. , 22(4), 2-7.
F Golchinfar; R Madani; GH.R Moazeni Jula. "Purification & separation of Bacillus anthracis 34F2 Antigens with UF & HPLC". , 22, 4, 2009, 2-7.
Golchinfar, F., Madani, R., Moazeni Jula, G. (2009). 'Purification & separation of Bacillus anthracis 34F2 Antigens with UF & HPLC', , 22(4), pp. 2-7.
Golchinfar, F., Madani, R., Moazeni Jula, G. Purification & separation of Bacillus anthracis 34F2 Antigens with UF & HPLC. , 2009; 22(4): 2-7.

Purification & separation of Bacillus anthracis 34F2 Antigens with UF & HPLC

تحقیقات دامپزشکی و فرآورده‌های بیولوژیک
Editorial, Volume 22, Issue 4, December 2010, Pages 2-7    PDF (443.38 K)
Authors
F Golchinfar* 1; R Madani2, 3; GH.R Moazeni Jula4
1,Biotechnology Dept. Razi Vaccine & Serum Research Institute
2Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
3Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4Aerobic Bacterial Vaccines Dept. Razi Vaccine & Serum Research Institute
Abstract
Anthrax is one of the important zoonose disease between human and cattle. Bacillus anthracis is a gram- positive bacterium which its tripartite protein toxin contains protective antigen (PA), edema factor (EF) and lethal factor (LF) and a poly D- glutamic acid capsule. A recent resurgence of interest in B.anthracis resulted in improved methods for production, separation and purification of the known toxin components. This in turn has lead to development of much more sensitive toxin-detection systems. Further understanding of toxin activity, the role of individual components in protection against the disease and the production of antigens for use in diagnostic requires high-degree of purity. In this study after cultivation of bacteria and separaing the supernatant the solution was passed through different membrane of  ultra filter system with 30000, 50000 and 100000 Dalton molecular weight cut off respectively. The final solution was treated with ammonium sulphate and the antigens prepared which after dialysis have passed through gel chromatography of HPLC for separating of  B.anthracis toxins. Finaly three toxin antigens EF,LF and PA  with 86, 87 and 85 K dalton molecular weight of high purity were obtained.
References

1. Bailie, L and Read, T.D. (2001) Bacillus anthracites, a bug with attitude. Curr opin  microbiol. 4:78-81##

2 - Quinn, C. P., Shone, C. C.,  Turnbull, P C, and Melling, J. (1988); Purification of anthrax-toxin components by high-performance anion-exchange, gel-filtration and hydrophobic-interaction chromatography. Biochem J.  15; 252(3): 753–758##

3-Chul-Soon, C., Sang-In, C., Ki-Jeong,K., Chan-Ho, C., Won-Yong, K and Yong-Tae, Y. (1996) Study on the antigens for cell free vaccine and retrospective diagnosis of anthrax: Production of  protective antigen of Bacillus anthracis. J.Korean Soc. Microbiol, 31(6), 631-641##

4. Doris, K and Larson ,P (1986) Separation of three exotoxic factors of bacillus anthracis by sequential immuno sorbent chromatography. J.Toxicon.26:10:913-921##

5.Farchaus, J.W and Ribot, W.J (1998) Fermentation, purification and characterization of protective antigen from a recombinant a virulent strain of Bacillus anthracis. Applied and environ microbial. 64:3: 982-991##

6. Gladstone, G.P (1946) Immunity to anthrax, protective antigen present in cell free culture filtration. Br.J.EXP.Pathol.27:394-418##

7. Heckly, R.I and Gold wasser, E (1949) Studies on identification Bacillus anthracis the production of an immunizing antigen invitro. I.Infect. 84-91##

8. Leppla, S.H.,  (1988) Production and purification of anthrax toxin. Meth.Eny.165-  168.##

9. Muder, S.H (1991) Background for after filtration of protein solutions. Cent.Afr.I.Med.42:312-15. ##

10.  Mabry,R.,  Mridula R., Geiger,R., Hubbard,G., Carrion, Jr R., Brasky,K.,  Patterson,J.L.,  Georgiou,G. and Iverson B. L(2005)Passive Protection against Anthrax by Using a High-Affinity Antitoxin  Antibody Fragment Lacking an Fc Region. J. Infection and Immunity.73(12) 8362-  8368##

11. Slack, R.C.B.(2002) Medical microbiology, a guide to infections: Pathogenesis, Immunity Laboratory diagnosis and control, 16nd Edn, CChurchill Lvingstone, London.##

12. Vincent, P and Guy, P. (1996) Identification and characterization of bacillus anthracis by multiplex PCR and pXO2 and chromosomal DNA. FEMS.Microbiol.9-16.##

13. Welkos, S.L. and Love. I.R. (1988) Sequenced and analysis of the DNA encoding protective antigen of Bacillus anthracis. Gene. 69:287-300.##  

Statistics
Article View: 1,265
PDF Download: 800


counter
Journal management system. designed by sinaweb