Publications (2)0 Total impact
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Article: [Preparation and identification of PON2 monoclonal antibodies.].
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ABSTRACT: To prepare and characterize the mouse monoclonal antibodies against human PON2 (paraoxonase 2). A fragment of human PON2 gene of low homology with mice but of strong hydrophilicity and immunogenicity was selected for recombinant expression. Mice were immunized with the purified HIS fusion protein 3 times. The specificity and sensitivity of the anti-human PON2 monoclonal antibodies were characterized by Western blot and indirect immunofluorescence. HIS-PON2 and GST-PON2 fusion protein was highly expressed in E.coli with a molecular weight of 40 kDa and 46 kDa. Western blot analysis proved that the established monoclonal antibodies could specifically recognize the target proteins expressed in E.coli expression system. Indirect immunofluorescence analysis confirmed that PON2 protein was located in the cytoplasm of HepG2 cells. The monoclonal antibodies against human PON2 can specifically recognize natural protein expressed in human cells, Which can be used for further functional study of PON2 protein.Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 12/2009; 25(12):1130-2. -
Article: [Expression of procalcitonin and characterization of antibodies against PCT].
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ABSTRACT: To construct the expression vectors of procalcitonin (PCT), prepare polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) against PCT and identify their specific biological activity. The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed using thyroid carcinoma cell line (TT cell) cDNA as template. The fusion protein of His-PCT was expressed in E.coli and used as immunogen. The specificity of antiserum against human PCT was characterized by ELISA, Western blot and indirect immunofluorescence. The mAbs against human PCT were identified by Western blot and indirect immunofluorescence. The recombinant expression plasmids of pGEX-4T-1-PCT and PET-32a-PCT were constructed and the fusion protein of His-PCT was expressed and purified. The antiserum against human PCT was prepared and the titer detected by ELISA was 1:256 000. The pAb specifically recognized the recombinant human PCT. Eight hybridoma cell lines secreting specific mAbs against PCT were established. The mAbs recognized the recombinant human PCT and four of them recognized the native PCT of TT cytoplasm in immunofluorescent assay. The successful preparation of polyclonal and monoclonal antibodies against human PCT is beneficial to further research into the pathological and physiological functions of PCT in severe bacterial infection and sepsis.Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 09/2008; 24(8):795-7.
