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[Screening and application of human-derived HBsAg bacteriophage single chain antibody in clinical diagnosis]
Yanwei Zhong1, Jun Cheng, Shuangshuang Shi
1Gene Therapy Research Centre, Institute of Infectious Diseases, The 302nd Hospital of PLA, Beijing 100039, China.
Objective:
To identify human single chain Fv antibody (ScFv) against hepatitis B viral surface antigen.
Methods:
The recombinant phages were panned by HBsAg which was coated in a microtiter plate, after five rounds of biopanning, 56 phage clones were identified specific to HBsAg. The specificity of ScFv was evaluated by ELISA and immunohistochemistry, respectively.
Results:
The data of HB sAg-ScFv DNA digestion and DNA sequencing showed that the ScFv gene is composed of 750 bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis B surface antigen has a specific combination character with hepatitis B surface antigen of different sources and paraffin-embedded patients tissue specimens, it did not react with normal liver tissue and HCV.
Conclusions:
The application of HBsAg specific ScFv in immunohistochemistry was successfully achieved.
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