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Golsaz Shirazi, Forough, Mehdi Amiri, Mohammad, Mohammadi, Hamed, Ahmad Bayat, Ali, Roohi, Azam, Khoshnoodi, Jalal, Zarnani, Amir Hassan, Jeddi-Tehrani, Mahmood, Kardar, Gholam Ali, Shokri, Fazel. (1392). Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis. سامانه مدیریت نشریات علمی, 10(3), 127-138.
Forough Golsaz Shirazi; Mohammad Mehdi Amiri; Hamed Mohammadi; Ali Ahmad Bayat; Azam Roohi; Jalal Khoshnoodi; Amir Hassan Zarnani; Mahmood Jeddi-Tehrani; Gholam Ali Kardar; Fazel Shokri. "Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis". سامانه مدیریت نشریات علمی, 10, 3, 1392, 127-138.
Golsaz Shirazi, Forough, Mehdi Amiri, Mohammad, Mohammadi, Hamed, Ahmad Bayat, Ali, Roohi, Azam, Khoshnoodi, Jalal, Zarnani, Amir Hassan, Jeddi-Tehrani, Mahmood, Kardar, Gholam Ali, Shokri, Fazel. (1392). 'Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis', سامانه مدیریت نشریات علمی, 10(3), pp. 127-138.
Golsaz Shirazi, Forough, Mehdi Amiri, Mohammad, Mohammadi, Hamed, Ahmad Bayat, Ali, Roohi, Azam, Khoshnoodi, Jalal, Zarnani, Amir Hassan, Jeddi-Tehrani, Mahmood, Kardar, Gholam Ali, Shokri, Fazel. Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis. سامانه مدیریت نشریات علمی, 1392; 10(3): 127-138.

Construction and Expression of Hepatitis B Surface Antigen Escape Variants within the "a" Determinant by Site Directed Mutagenesis

Iranian Journal of Immunology
مقاله 1، دوره 10، شماره 3، آذر 2013، صفحه 127-138    اصل مقاله (376.26 K)
نوع مقاله: Original Article
نویسندگان
Forough Golsaz Shirazi1؛ Mohammad Mehdi Amiri1؛ Hamed Mohammadi1؛ Ali Ahmad Bayat2؛ Azam Roohi1، 3؛ Jalal Khoshnoodi1؛ Amir Hassan Zarnani4؛ Mahmood Jeddi-Tehrani2؛ Gholam Ali Kardar5؛ Fazel Shokri* 1، 2
1Department of Immunology, School of Public Health, Tehran University of Medical Sciences
2Monoclonal Antibody Research Center
3Department of Molecular Medicine, School of Advanced Medical Technologies, Tehran University of Medical Sciences
4Reproductive Immunology Research Center, Avicenna Research Institute, ACECR
5Immunology, Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran
چکیده
Background: The antibody response to hepatitis B surface antigen (HBsAg) controls hepatitis B virus infection. The "a" determinant of HBsAg is the most important target for protective antibody response, diagnosis and immunoprophylaxis. Mutations in this area may induce immune escape mutants and affect the performance of HBsAg assays.
Objectives: To construct clinically relevant recombinant mutant forms of HBsAg and assessment of their reactivity with anti-HBs monoclonal antibodies (MAbs).
Methods: Wild type (wt) and mutant (mt) HBsAg genes were constructed by site directed mutagenesis and SEOing PCR. The amplified genes were inserted into pCMV6-neo plasmid and transfected in CHO cell line. The expression of wt- and mtHBsAg was assessed by commercial ELISA assays and stable cells were established and cloned by limiting dilution. The recombinant mutants were further characterized using a panel of anti-HBs monoclonal antibodies (MAbs) and the pattern of their reactivity was assessed by ELISA.
Results: Ten HBsAg mutants having single mutation within the "a" determinant including P120E, T123N, Q129H, M133L, K141E, P142S, D144A, G145R, N146S and C147S together with a wt form were successfully constructed and expressed in CHO cells. Reactivity of anti-HBs MAbs with mtHBsAgs displayed different patterns. The effect of mutations on antibody binding differed depending on the amino acid involved and its location within the ‘‘a’’ determinant. Mutation at amino acids 123 and 145 resulted in either complete loss or significant reduction of binding to all anti-HBs MAbs.
Conclusion: Our panel of mtHBsAgs is a valuable tool for assessment of the antibody response to HBV escape mutants and may have substantial implications in HBV immunological diagnostics.
کلیدواژه‌ها
"a" Determinant؛ HBs Ag؛ Monoclonal antibody
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