EXTRACTION, PURIFICATION AND DETOXIFICATION OF BRUCELLA ABORTUS LIPOPOLYSACCHARIDE AND BIOLOGICAL ACTIVITIES EVALUATION

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PAKZAD I.; REZAEI A.; RASAEI M.J.; ZAVARAN HOSSEINI A.; KAZEMNEJAD ANOUSHIRAVAN; TABARAEI BAHMAN; RAIVANDI S.

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Journal: Hakim Journal Year:2004 | Volume:6 | Issue:4 Page(s): 73-79

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Title

EXTRACTION, PURIFICATION AND DETOXIFICATION OF BRUCELLA ABORTUS LIPOPOLYSACCHARIDE AND BIOLOGICAL ACTIVITIES EVALUATION

Author(s)

Keywords

LIPOPOLYSACCHARIDE (LPS)Q4

BRUCELLA ABORTUSQ4

DETOXIFICATIONQ4

LALQ4

RABBIT PYROGENIC TESTQ4

Abstract

Introduction: Lipopolysaccharide (LPS) of BRUCELLA ABORTUS is important in brucellosis diagnosis and as one of the components for developing a subunit vaccine against brucellosis. Comparision of LPS extraction methods and evaluation of B. abortus LPS toxicity have been considered as the goal of this research.Methods: BRUCELLA ABORTUS LPS was extracted by n-butanol and hot phenol method, in E.coli by hot phenol, then they were purified by ultracentrifugation. B. abortus LPS was detoxified by alkaline treatment. LPS SDS-PAGE was performed with polyacrylamide gels and stained with silver stain. Results: Purified LPS from B.abortus by butanol extraction was shown to have less than 2% (wt/wt) contamination by protein and less than 1%(wt/wt) contamination by nucleic acids. In addition, purified LPS from B.abortus and E.coli extracted by phenol were shown to have below 4% (wt/wt) contamination by protein and below 1.5% contamination by nucleic acids. The electrophoresis pattern of B.abortus LPS and E.coli LPS was the same as the former studies. Pyrogenicity test of B.abortus LPS (10 μg/ml) and E.coli LPS (0.5 μg/ml) was positive, but D-LPS (10, 50μg/ml) was negative. In LAL test, 10 ng/ml of D-LPS was negative, but 0.04ng/ml of B.abortus LPS was positive and endotoxin unit of B.abortus LPS was less than E.coli LPS.Conclusion: Results show that the amount of LPS of extracted by phenol method is higher, and protein, nucleic acids contamination rate in butanol method is very low. In addition, D-LPS toxicity is severely decreased, hence, we can use several as many as B.abortus LPS for stimulating immune system. Besides, ability of B.abortus LPS is likely much less than the LPS from E.coli to evoke endotoxic shock, and it can be used as immunogen directly.

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APA: Copy

PAKZAD, I., REZAEI, A., RASAEI, M.J., ZAVARAN HOSSEINI, A., KAZEMNEJAD, ANOUSHIRAVAN, TABARAEI, BAHMAN, & RAIVANDI, S.. (2004). EXTRACTION, PURIFICATION AND DETOXIFICATION OF BRUCELLA ABORTUS LIPOPOLYSACCHARIDE AND BIOLOGICAL ACTIVITIES EVALUATION. HAKIM RESEARCH JOURNAL, 6(4), 73-79. SID. https://sid.ir/paper/357043/en

Vancouver: Copy

PAKZAD I., REZAEI A., RASAEI M.J., ZAVARAN HOSSEINI A., KAZEMNEJAD ANOUSHIRAVAN, TABARAEI BAHMAN, RAIVANDI S.. EXTRACTION, PURIFICATION AND DETOXIFICATION OF BRUCELLA ABORTUS LIPOPOLYSACCHARIDE AND BIOLOGICAL ACTIVITIES EVALUATION. HAKIM RESEARCH JOURNAL[Internet]. 2004;6(4):73-79. Available from: https://sid.ir/paper/357043/en

IEEE: Copy

I. PAKZAD, A. REZAEI, M.J. RASAEI, A. ZAVARAN HOSSEINI, ANOUSHIRAVAN KAZEMNEJAD, BAHMAN TABARAEI, and S. RAIVANDI, “EXTRACTION, PURIFICATION AND DETOXIFICATION OF BRUCELLA ABORTUS LIPOPOLYSACCHARIDE AND BIOLOGICAL ACTIVITIES EVALUATION,” HAKIM RESEARCH JOURNAL, vol. 6, no. 4, pp. 73–79, 2004, [Online]. Available: https://sid.ir/paper/357043/en

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