ABSTRACT
In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E. coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 x 10(9) cfu/microg was obtained. After amplification with helper phage, the titer of antibody library reached 5 x 10(12) cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.
ABSTRACT
In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E.coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 × 109 cfu/μg was obtained. After amplification with helper phage, the titer of antibody library reached 5 × 1012 cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.
ABSTRACT
Summary: Receptor mediated gene delivery is a new gene transfer strategy. Asialoglycoprotein receptor (ASGP-R), the receptor of asialoorosomucoid (Asor), is specially expressed on the surface of hepatocyte. In this paper, the nuclide 131I was combined with Asor to form a kind of soluble nuclide-protein complex, which can be specifically endocytosed into hepatocyte by ASGP-R. After intravenous injection of the complex into experimental animals, the deposition of Asor in vivo and the targeting quality of hepatocyte was detected by ECT. This research testified the feasibility of targeting Asor complex delivery to hepatocyte mediated by ASGP-R in vivo, and provided foundation for the genetic diagnosis and gene therapy of hepatic cell-related diseases.
ABSTRACT
To explore the possibility to employ 99mTc-MIBI to monitor biological response of tumor cells after irradiation and to observe the relation between the radiation doses and the uptake levels of 99mTc-MIBI in tumor cells, the cells were irradiated with a single dose of 2 Gy, 10 Gy and 20 Gy respectively. The uptake of 99mTc-MIBI in each dosage group was determined before and 24, 48, 72 h after irradiation respectively. Apoptosis index (AI), plating efficiency (PE) of tumor cells was simultaneously determined. There was a positive correlation between uptake levels of 99mTc-MIBI and AI(r=－0.91, P<0.05). A negative correlation was noted between the uptake levels and PE (r=－0.86, P<0.05). It is suggested that 99mTc-MIBI may be used as a tracer to monitor the change of viability state of tumor cells after being irradiated with different doses.
ABSTRACT
To explore the possibility to employ 99mTc-MIBI to monitor biological response of tumor cells after irradiation and to observe the relation between the radiation doses and the uptake levels of 99mTc-MIBI in tumor cells, the cells were irradiated with a single dose of 2 Gy, 10 Gy and 20 Gy respectively. The uptake of 99mTc-MIBI in each dosage group was determined before and 24, 48, 72 h after irradiation respectively. Apoptosis index (AI), plating efficiency (PE) of tumor cells was simultaneously determined. There was a positive correlation between uptake levels of 99mTc-MIBI and AI(r=－0.91, P<0.05). A negative correlation was noted between the uptake levels and PE (r=－0.86, P<0.05). It is suggested that 99mTc-MIBI may be used as a tracer to monitor the change of viability state of tumor cells after being irradiated with different doses.