[Expression and bioactivity analysis of the fusion protein GFP-V(L) of the neutralizing monoclonal antibody MA18/7 against HBV].
نویسندگان
چکیده
AIM To express the fusion protein of enhanced green fluorescent protein (EGFP) with the light chain variable domain of the neutralizing monoclonal antibody MA18/7 (mAb) against hepatitis B virus in E.coli, and determine its bioactivity. METHODS The EGFP gene was cloned into vector pTO-T7 to construct an expression vector. And then according to ORF gene, MA18/7-V(L) was inserted into the 5' terminal of EGFP gene free of terminal code TAA to construct expression vector of fusion protein. The fusion protein was expressed in E.coli and its bioactivity was detected with ELISA and relative fluorescence intensity. RESULTS The expression vector EGFP-V(L) was constructed. SDS-PAGE analysis showed that expressed fusion protein was mainly in the form of inclusion body. The fusion protein retained the property of EGFP and it could bind to V(H) to form Fv which had binding activity to pre-S1. CONCLUSION The obtained fusion protein had good bioactivity and could be applied to further studies.
منابع مشابه
Production of monoclonal antibody against recombinant NS3 protein of bovine viral diarrhea virus (NADL strain)
Bovine Viral Diarrhea virus (BVDV) is an important viral pathogen of cattle causing several clinical syndromes. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on serological detection and virus isolation. Nonstructural protein 3 (NS3) as immunogenic protein of BVDV is genetically and antigenically conserved among different isolates....
متن کاملProduction and Characterization of a Monoclonal Antibody Neutralizing Poliovirus Type 2
Monoclonal Antibodies (mAbs) are used for biomedical research, diagnosis, treatment, production, and the quality control of biological products. mAbs are also very helpful in poliovirus research studies because it is still one of the major public health problems in developing countries. The main objective of this study was the production of mAbs against Poliovirus Type 2 (PV2) to be prepared an...
متن کاملDesign and Construction of a Novel Humanized Single-Chain Variable-Fragment Antibody against the Tumor Necrosis Factor alpha
The pro-inflammatory cytokine, TNF-α, which plays a major role in the development and persistence of inflammatory diseases, is the basis for the use of anti-TNF-α therapies. The neutralization of TNF-α or blockage of its binding to the corresponding receptor has mainly served as a therapeutic strategy against some diseases. This study aimed to investigate the production of a humanized single ch...
متن کاملDesign and Construction of a Novel Humanized Single-Chain Variable-Fragment Antibody against the Tumor Necrosis Factor alpha
The pro-inflammatory cytokine, TNF-α, which plays a major role in the development and persistence of inflammatory diseases, is the basis for the use of anti-TNF-α therapies. The neutralization of TNF-α or blockage of its binding to the corresponding receptor has mainly served as a therapeutic strategy against some diseases. This study aimed to investigate the production of a humanized single ch...
متن کاملProduction of Monoclonal Antibody against Prokaryotically Expressed G1 Protein of Bovine Ephemeral Fever Virus
Epitope-G1 of bovine ephemeral fever virus (BEFV) G glycoprotein has been genetically and antigenically conserved among various isolates of BEFV and only reacts with anti-BEFV neutralising antibodies. Therefore, it is a candidate antigen for development of the enzyme linked immunosorbent assay (ELISA) for serological identification bovine ephemeral fever (BEF)-infected animals. The aim of this ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
دوره 20 4 شماره
صفحات -
تاریخ انتشار 2004