EXPRESSION AND PURIFICATION OF NEUROTOXIN-ASSOCIATED PROTEIN HA-33/A FROM CLOSTRIDIUM BOTULINUM AND EVALUATION OF ITS ANTIGENICITY

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SAYADMANESH ALI; EBRAHIMI FIROUZ; HAJIZADEH ABBAS; ROSTAMIAN MOSAYEB; KESHAVARZ HANI

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Journal Paper

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Journal: Iranian Biomedical Journal Year:2013 | Volume:17 | Issue:4 Page(s): 165-170

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Title

EXPRESSION AND PURIFICATION OF NEUROTOXIN-ASSOCIATED PROTEIN HA-33/A FROM CLOSTRIDIUM BOTULINUM AND EVALUATION OF ITS ANTIGENICITY

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Keywords

BOTULINUM NEUROTOXIN

EXPRESSION

PURIFICATION

Abstract

Background: BOTULINUM NEUROTOXIN (BoNT) complexes consist of neurotoxin and neurotoxin-associated proteins.Hemagglutinin-33 (HA-33) is a member of BoNT type A (BoNT/A) complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expressed and purified, and subsequently its antigenicity in mice was studied.Methods: Initially, ha-33 gene sequence of Clostridium botulinum serotype A was adopted from GenBank. The gene sequence was optimized and synthesized in pET28a (+) vector. E. coli BL21 (DE3) strain was transformed by the recombinant vector and the EXPRESSION of HA-33 was optimized at 37oC and 5 h induction time.Results: The recombinant protein was purified by nickel nitrilotriacetic acid agarose affinity chromatography and confirmed by immunoblotting. Enzyme Linked Immunoassay showed a high titer antibody production in mice.Conclusion: The results indicated a highly expressed and purified recombinant protein, which is able to evoke high antibody titers in mice.

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APA: Copy

SAYADMANESH, ALI, EBRAHIMI, FIROUZ, HAJIZADEH, ABBAS, ROSTAMIAN, MOSAYEB, & KESHAVARZ, HANI. (2013). EXPRESSION AND PURIFICATION OF NEUROTOXIN-ASSOCIATED PROTEIN HA-33/A FROM CLOSTRIDIUM BOTULINUM AND EVALUATION OF ITS ANTIGENICITY. IRANIAN BIOMEDICAL JOURNAL, 17(4), 165-170. SID. https://sid.ir/paper/277361/en

Vancouver: Copy

SAYADMANESH ALI, EBRAHIMI FIROUZ, HAJIZADEH ABBAS, ROSTAMIAN MOSAYEB, KESHAVARZ HANI. EXPRESSION AND PURIFICATION OF NEUROTOXIN-ASSOCIATED PROTEIN HA-33/A FROM CLOSTRIDIUM BOTULINUM AND EVALUATION OF ITS ANTIGENICITY. IRANIAN BIOMEDICAL JOURNAL[Internet]. 2013;17(4):165-170. Available from: https://sid.ir/paper/277361/en

IEEE: Copy

ALI SAYADMANESH, FIROUZ EBRAHIMI, ABBAS HAJIZADEH, MOSAYEB ROSTAMIAN, and HANI KESHAVARZ, “EXPRESSION AND PURIFICATION OF NEUROTOXIN-ASSOCIATED PROTEIN HA-33/A FROM CLOSTRIDIUM BOTULINUM AND EVALUATION OF ITS ANTIGENICITY,” IRANIAN BIOMEDICAL JOURNAL, vol. 17, no. 4, pp. 165–170, 2013, [Online]. Available: https://sid.ir/paper/277361/en

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