Production and Characterization of Murine Monoclonal Antibodies Recognizing Conformational and Linear Epitopes Localized on Human IgA2 Molecules

نویسندگان

  • Fatemeh Hajighasemi Department of Immunology, School of Public Health, Tehran University of Medical Sciences | Department of Immunology, School of Medicine, Shahed University
  • Fazel Shokri Department of Immunology, School of Public Health, Tehran University of Medical Sciences | Monoclonal Antibody Research Center, Avesina Research Center, Tehran, Iran
  • Jalal Khoshnoodi Department of Immunology, School of Public Health, Tehran University of Medical Sciences
  • Nasrin Moheghi Department of Immunology, School of Public Health, Tehran University of Medical Sciences
  • Roya Ghods Monoclonal Antibody Research Center, Avesina Research Center, Tehran, Iran
  • Soheila Gharagozlou Department of Immunology, School of Public Health, Tehran University of Medical Sciences
چکیده مقاله:

Background: There are two subclasses of human IgA (IgA1 and IgA2) that differ in antigenic properties and in chemical composition. The constant domains of α1 and α2 heavy chains have >95% sequence homology though major structural differences exist in the hinge region. Quantitation of IgA subclass levels depends on the availability of monoclonal antibodies (MAbs) specific for conserved conformational or linear epitopes restricted to each subclass. Objective: To produce, select and characterize monoclonal antibodies specific for human IgA2. Methods: Splenocytes from BALB/C mice immunized with a human IgA2 myeloma protein were fused with SP2/0 myeloma cells. Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by limiting dilution assay. Antibody (Ab) secreting cells were screened by enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAbs was further analyzed, using a panel of purified myeloma proteins and some animal sera by ELISA and immunoblotting. The affinity constant (Kaff) was also determined by ELISA. Results: Four murine hybridoma clones designated 2F20G5, 2F20B5, 3F20E3 and 6F20H11 were obtained that secreted MAbs specific for the human IgA2. 2F20G5 and 6F20H11 MAbs react with linear epitope(s) while 2F20B5 and 3F20E3 react with conformational epitope(s) located to human IgA2 subclass. 2F20G5 MAb displays a weak cross-reactivity with monkey and rabbit sera and a strong cross-reactivity with cat and dog sera while the other three MAbs showed no cross-reactivity with the animal sera tested. Conclusion: These MAbs, especially 6F20H11 with high affinity constant (6.03 ×109 M-1) are useful tools for quantitation of human IgA2 subclass levels in various diseases. Cross-reactivity of 2F20G5 MAb with some animalsera suggests phylogenic conservation ofthe epitope recognized by this MAb.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

production and characterization of murine monoclonal antibodies recognizing conformational and linear epitopes localized on human iga2 molecules

background: there are two subclasses of human iga (iga1 and iga2) that differ in antigenic properties and in chemical composition. the constant domains of α1 and α2 heavy chains have >95% sequence homology though major structural differences exist in the hinge region. quantitation of iga subclass levels depends on the availability of monoclonal antibodies (mabs) specific for conserved conformat...

متن کامل

production and characterization of mouse monoclonal antibodies recognizing human pan-igg specific conformational or linear epitopes

background: pan-igg specific monoclonal antibodies (mabs) are essential tools for assessment of humoral immunity, immune deficiency and gammopathy. in this study, four hybridoma clones producing mabs with different specificities for human igg isotypes were established. methods: splenocytes from balb/c mice immunized with fc fractions of human igg were fused with sp2/0 myeloma cells. hybridoma c...

متن کامل

Production and Characterization of Mouse Monoclonal Antibodies Recognizing Human Pan-IgG Specific Conformational or Linear Epitopes

BACKGROUND Pan-IgG specific monoclonal antibodies (MAbs) are essential tools for assessment of humoral immunity, immune deficiency and gammopathy. In this study, four hybridoma clones producing MAbs with different specificities for human IgG isotypes were established. METHODS Splenocytes from Balb/c mice immunized with Fc fractions of human IgG were fused with SP2/0 myeloma cells. Hybridoma c...

متن کامل

Production and Characterization of Monoclonal Antibodies Recognizing a Common 57-kDa Antigen of Leishmania Species

Background: The therapy of leishmania infection is difficult and each year 1.5 million new cases of cutaneous leishmaniasis and 500,000 new cases of visceral leishmaniasis are estimated, therefore, there is a need for an effective vaccine. Monoclonal antibody (mAb) is one of the suitable methods for isolation and purification of leishmania antigens. In this report, we produced several mAb aga...

متن کامل

Production and Characterization of Murine Monoclonal Antibodies to Leishmania Gp63 Antigen

Background : Production of monoclonal antibodies to Leishmania antigens assists the identification and characterization of these organisms. Objective: Production of monoclonal antibodies against epitopes on the gp63. Methods: Two murine monoclonal antibodies to gp63 were produced and characterized. The reactions of both antibodies with soluble leishmanial antigens, purified gp63 and truncated r...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

ذخیره در منابع من قبلا به منابع من ذحیره شده

{@ msg_add @}


عنوان ژورنال

دوره 2  شماره 3

صفحات  125- 133

تاریخ انتشار 2005-09-01

با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023