• 대한방사성의약품학회지
• /
• 제3권2호
• /
• pp.54-58
• /
• 2017

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) is one of the powerful methods that enable analysis of small molecules as well as large molecules up to about 500,000 Da without severe fragmentation. MALDI-TOF MS, thus, has been a very useful an analytical tool for the confirmation of synthetic molecules, probing PTMs, and identifying structures of a given protein. In recent nuclear medicine, MALDI-TOF MS liner ion mode helps researcher calculate the average number of chelator(or linkage) per an antibody conjugate, such as DOTA-(or DFO-) trastuzumab for labeling a medical radioisotope. This simple technique can be utilized to improve the labeling method and control the quality at the development of antibody-based radiopharmaceuticals, which is very effected to diagnosis and therapy for in vivo tumor cells, with radioisotopes like $^{89}Zr$, $^{64}Cu$, and 177Lu. To minimize the error, MALDI-TOF MS measurement is repeatedly performed for each sample in this study, and external calibration is carried out after data collection.

• Radiation Oncology Journal
• /
• 제15권3호
• /
• pp.225-231
• /
• 1997

목적 : 두경부암 환자에서 수술 및 수술 후 방사선치료가 갑상선 기능에 미치는 영향을 평가하기 위하여 전향적 연구를 시행하였다. 대상 및 방법 : 1986년 9월부터 1994년 12월까지 수술후 방사선치료를 받은 두경부암 환자 중 추적 관찰이 가능했던 71예를 대상으로 하였다. 환자의 연령분포는 32세부터 73세였고, 중간값은 58세였으며, 여자 12예, 남자 59예였다. 원발 병소부위는 후두가 34예로 가장 많았고, 구강 12예, 구인두 1예, 하인두 13예, 상악동 2예, 타액선 3예, 근원 불명 전이성 경부암 6예였다. 전후두절 제술 및 경부곽청수술을 시행한 환자는 45예였고, 경부곽청수술만 시행한 환자는 26예였다. 모든 환자에 대하여 방사선치료 전 및 방사선치료 후에 정기적으로 임상검사 및 갑상선 기능검사 (T3, T4, free T4, TSH, antithyroglovulin antibody, antimicrosomal antibody)를 시행하였다. 방사선량은 40.6Gy에서 60Gy였고 중간값은 50Gy였으며 추적관찰 기간은 3개월에서 80개월이었다. 결과 : 갑상선 기능 이상의 빈도는 $56.3\%(40/71)$였다. 31예 $(43.7\%)$는 갑상선 기능이 정상이었고 7예 $(9.9\%)$는 임상적 갑상선 기능저하증이었으며 33예 $(46.5\%)$는 준임상적 갑상선 기능저하증이었다. 갑상선 기능항진증, 갑상선 결절, 악성종양은 발견되지 않았다. 경부곽청수술군보다 전후두절제술 및 경부곽청수술군에서 갑상선 기능 이상이 일찍 발생하였다(p<0.05). 방사선치료 후 갑상선 기능 이상의 발생 빈도에 영향을 준 위험인자는 전후두절제술 및 경부곽청수술과 방사선치료의 병용요법 (P=0.0000)이었다. 전후두절제술 및 경부곽청수술군 45예 중 36예 $(80\%)$에서 갑상선 기능 이상이 발생하였으며 경부곽청수술군 26예 중 4예 $(15.4\%)$에서만이 갑상선 기능이상이 발생하였다. 결론 : 두경부암 환자에서 수술 및 방사선치료 후 갑상선 기능 이상은 비교적 흔한 합병증이며, 갑상선 기능 이상의 발생 빈도에 영향을 미치는 인자는 전후두절제술 및 경부곽청수술과 방사선치료의 병용요법이었다. 갑상선 기능 이상을 조기에 발견하여 치료하기 위하여 방사선치료 전 및 방사선치료 후에 정기적인 갑상선 기능검사가 필요할 것으로 사료된다.

• Parasites, Hosts and Diseases
• /
• 제54권1호
• /
• pp.93-96
• /
• 2016

Toxoplasmosis, a neglected tropical disease caused by the protozoan parasite Toxoplasma gondii, occurs throughout the world. Human T. gondii infection is asymptomatic in 80% of the population; however, the infection is life-threatening and causes substantial neurologic damage in immunocompromised patients such as HIV-infected persons. The major purpose of this study was to investigate the seroprevalence of T. gondii infection in subjects infected with HIV/AIDS in eastern China. Our findings showed 9.7% prevalence of anti-T. gondii IgG antibody in HIV/AIDS patients, which was higher than in intravenous drug users (2.2%) and healthy controls (4.7%), while no significant difference was observed in the seroprevalence of anti-Toxoplasma IgM antibody among all participants (P>0.05). Among all HIV/AIDS patients, 15 men (7.7%) and 10 women (15.9%) were positive for anti-T. gondii IgG antibody; however, no significant difference was detected in the seroprevalence of anti-Toxoplasma IgG antibody between males and females. The frequency of anti-Toxoplasma IgG antibody was 8.0%, 13.2%, 5.5%, and 0% in patients with normal immune function ($CD4^+$ T-lymphocyte count ${\geq}500cells/ml$), immunocompromised patients (cell count ${\geq}200$ and <500 cells/ml), severely immunocompromised patients (cell count ${\geq}50$ and <200 cells/ml), and advanced AIDS patients, respectively (cell count <50 cells/ml), while only 3 immunocompromised patients were positive for anti-T. gondii IgM antibody. The results indicate a high seroprevalence of T. gondii infection in HIV/AIDS patients in eastern China, and a preventive therapy for toxoplasmosis may be given to HIV/AIDS patients based on $CD4^+$ T lymphocyte count.

• 대한핵의학회:학술대회논문집
• /
• 대한핵의학회 2006년도 제45차 춘계학술대회
• /
• pp.25-25
• /
• 2006

• IMMUNE NETWORK
• /
• 제5권1호
• /
• pp.11-15
• /
• 2005

Background: Throughtout the last three decades, the therapy of leukemias and lymphoma has set the stage for curative cancer therapy in systemic malignant disease. This was the result of an integrated work of basic reaserch and clinical investigators leading to more aggressive albeit tolerable protocol of chemotherapy and radiotherapy. High dose therapy marks the most elaborated strategies in this field today. However, intensification of conventional therapeutic modalities as mentioned has to be based on new approaches and the exploration of new antineoplastic mechanisms. This insight has resulted in immune therapy of cancer. Among the cells of the immune system, natural killer (NK) cells and T cells are of major interest for the development of therapeutic strategies. Methods: Cytotoxicity to target cells was measured by LDH release method, Characterization of activated lymphocyte was measured by Flow cytometry analysis. Anti-CD3, 16, 56 monoclonal antibody and IL-2 were used for the activation of NK and T cell. The analysis of effect of activated lymphocyte, in vivo, were used by Balb/c nude mouse. Results and Conclusion: Cytotoxicity to K562 cells was significantly higher in the mixture group of NK and T cells than that of a group of activating T cells. The survivors and the rate of reduction of size of tumor craft of nude mouse group treatment with activated lymphocyte was higher than that of the group without treatment with activated lymphocyte. Therefore, this results are suggested that the activated lymphocytes by anti-CD3, CD16 and CD56 can reduce the malignancy effect of lymphoma.

• 대한핵의학회지
• /
• 제29권4호
• /
• pp.451-459
• /
• 1995

Thirty-eight patients with metastatic well-differentiated thyroid carcinoma treated with 200mCi $^{131}I$ were studied. There were false negative serum thyroglobulin values during TSH suppression or at anti-thyroglobulin antibody(+) and discrepancies between findings of whole body scan and serum thyroglobulin level. After one to five cycles of 200mCi $^{131}I$ therapy, complete remission and partial remission were achieved at 5.3% and 57.9%, respectively. We concluded that all of serum thyroglobulin, TSH, anti-thyroglobulin antibody, $^{131}I$ or $^{123}I$ whole body scan were necessary in follow up of metastatic well-differentiated thyroid carcinoma. Also, if there was no response after repetitive 200mCi $^{131}I$ therapy, higher doses of $^{131}I$ therapy should be considered.

• BMB Reports
• /
• 제42권11호
• /
• pp.731-736
• /
• 2009

The generation of functional recombinant antibodies from hybridomas is necessary for antibody engineering. However, this is not easily accomplished due to high levels of aberrant heavy and light chain mRNAs, which require a highly selective technology that has proven complicated and difficult to operate. Herein, we attempt to use an alkaline phosphate (AP)-fused form of single-chain variable fragment (scFv) for the simple identification of a hybridoma-derived, functional recombinant antibody. As a representative example, we cloned the scFv gene from a hybridoma-producing mouse IgG against branched-chain keto acid dehydrogenase complex-E2 (BCKD-E2) into an expression vector containing an in-frame phoA gene. Functional recombinant antibodies were easily identified by conventional enzyme-linked immunosorbent assay (ELISA) by employing scFv-AP fusion protein, which also readily serves as a valuable immuno-detective reagent.

• IMMUNE NETWORK
• /
• 제4권1호
• /
• pp.7-15
• /
• 2004

Background: The heat shock proteins (HSPs) play an important role in cellular protection mechanisms against physical or chemical stresses. In this study scFv antibodies specific for human HSP70.1 were isolated from a semi-synthetic human scFv library with the ultimate goal of developing anti-HSP70.1 intracellular antibody (intrabody) that may offer an attractive alternative to gene targeting to study the function of the protein in cells. Methods: A semi-synthetic human scFv display library ($5{\times}10^{8}$ size) was constructed using pCANTAB-5E vector and the selection of the library against bacterially expressed recombinant human HSP70.1 was attempted by panning. Results: Three positive clones specific for recombinant HSP70.1 were identified. All three clones used $V_{H}$ subgroup III. On the other hand, $V_{L}$ of two clones belonged to the kappa light chain subgroup I, but the other utilized $V_{k}$ subgroup IV Interestingly, these scFv molecules specifically reacted to the recombinant HSP70.1, yet failed to recognize native HSP70 induced in U937 human monocytic cells by heat treatment. Conclusion: Our results indicated that affinity selection of an scFv phage display library using recombinant antigens produced in E. coli might not guarantee the isolation of scFv antibody molecules specific for a native form of the antigen. Therefore, the source of target antigens needs to be chosen carefully in order to isolate biofunctional antibody molecules.

• IMMUNE NETWORK
• /
• 제1권1호
• /
• pp.7-13
• /
• 2001

Currently 18 monoclonal antibodies were approved by FDA for inj ection into humans for therapeutic or diagnostic purpose. And 146 clinical trials are under way to evaluate the efficacy of monoclonal antibodies as anti-cancer agents, which comprise 9 % of clinical trials in cancer therapy field. When considering a lot of disappointment and worries existed in this field during the past 15 years, this boom could be called as resurrection. Antibodies have several merits over small molecule drug. First of all it is easier and faster in development, as proper immunization of the target proteins usually raises good antibody response. The side effects of antibodies are more likely to be checked out in immunohistomchemical staining of whole human tissues. Antibody has better pharmacokinetics, which means a longer half-life. And it is non-toxic as it is purely a "natural drug. Vast array of methods was developed to get the recombinant antibodies to be used as drug. The mice with human immunoglobulin genes were generated. Fully human antibodies can be developed in fast and easy way from these mice through immunization. These mice could make even human monoclonal antibodies against any human antigen like albumin. The concept of combinatorial library was also actively adopted for this purpose. Specific antibodies can be screened out from phage, mRNA, ribosomal library displaying recombinant antibodies like single chain Fvs or Fabs. Then the coding genes of these specific antibodies are obtained from the selected protein-gene units, and used for industrial scale production. Both $na\ddot{i}ve$ and immunized libraries are proved to be effective for this purpose. In post-map arena, antibodies are receiving another spotlight as molecular probes against numerous targets screened out from functional genomics or proteomics. Actually many of these antibodies used for this purpose are already human ones. Through alliance of these two actively growing research areas, antibody would play a central role in target discovery and drug development.

• BMB Reports
• /
• 제44권4호
• /
• pp.244-249
• /
• 2011

The quality of a phage-displayed antibody library deteriorates with clonal variations, which are caused by differentially expressed Escherichia coli antibody genes. Using the human Fab SP114 against the pyruvate dehydrogenase complex-E2 (PDCE2), we created four E. coli TOP10F' clones with a pCMTG phagemid encoding Fab-pIII (pCMTG-Fab), Fd ($V_H+C_{H1}$)-pIII (pCMTG-Fd), or light chain (L) (pCMTG-L), or the vector only (pCMTG-${\Delta}Fab$) to investigate the effect of clonal variations in a defined manner. Compared to the others, the E. coli clone with pCMTG-Fab was growth retarded in liquid culture, but efficiently produced phage progenies by Ex12 helper phage superinfection. Our results suggest that an antibody library must be cultured for a short duration before helper phage superinfection, and that the Ex12 helper phage helped to alleviate the detrimental effect of clonal variation, at least in part, by preferentially increasing functional phage antibodies during phage amplification.