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Generation of 1D4 Single Chain Fv-Fc
홍권표,문유리,지형진,안병우,송형근 충북대학교 동물의학연구소 2011 Journal of Biomedical and Translational Research Vol.12 No.3
Immunotherapeutic approaches using agonist antibodies or fusion proteins of immunomodulatory molecules significantly inhibit tumor growth and boost cell-mediated immunity. We isolated mRNA from previously reported 1D4 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reverse-transcriptase polymerase chain reaction. Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1D4 single-chain Fv-Fc (scFv-Fc) constructs. A549 cells were used to present the 1D4 antigen. The secreted 1D4 scFv-Fc constructs were analyzed by flow cytometry. The DNA sequence of 1D4 scFv-Fc was obtained. The 1D4 scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was lower than the murine 1D4 antibody for A549 cells. A 1D4 scFv-Fc construct for immunotherapy was developed.
0.412 MeV 감마선에 대한 원주형 NaI(Tl) 섬광체의 총 절대검출효율 계산
홍권표,신희성,이상윤,노성기 대한방사선 방어학회 2002 방사선방어학회지 Vol.27 No.4
Total absolute detection efficiencies of a 7.62 cm(dia.) and 7.62 cm(height) cylindrical NaI(Tl) crystal have been calculated for 0.412 MeV r -rays from a source(point, circular disk, square and line type). In this calculation the linear energy-absorption coefficients based on Hubbell's data have been considered and then calculated total absolute detection efficiencies compared with those from Grosjean and Bossaert. Besides, the source axis-to-detector axis shift distance which, could give rise to about 0.05% deviation in the total absolute detection efficiencies has been calculated for a line-type source of 0.5 cm in its length when a source-to-detector distance is 5 cm. It is revealed that the total absolute detection efficiencies obtained in this study are considerably different from those of Grosjean and Bossaert. In addition it is found that the deviation induced due to an imperfect center of a line type source may be within 0.05% if the shifted discrepancy is no larger than 1.74mm.