• 대한세포병리학회지
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• 제15권1호
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• pp.17-27
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• 2004

To investigate a population-based survey of the prevalence of human papillomavirus (HPV) infection in South Korea, we performed Papanicolaou smears and tests for HPV DNA and anti-HPV antibody detection in 909 sexually active general women (age range; 20-74 years, median 44 years) who were randomly selected residents from S district of Busan City. The presence of DNA of 36 different HPV types was detected by means of a GP 5+/6+ primer-mediated PCR enzyme immunoassay in cervical exfoliated cells, and IgG antibodies against L1 virus-like particles (anti-VLPs) of 5 HPV types 16, 18, 31, 33, and 58 were tested by means of enzyme linked immunoassay. The incidence of cytologic abnormality was 5.2% in Pap smear. The positive rate of HPV DNA was 10.4%, high in young women younger than 35 years old and proportionally increased according to the cytologic grades. The most often found HPV type was HPV 70, followed by HPV 16 and 33, and high-risk HPV types were more frequent in women younger than 35 years old. The most common HPV type in abnormal cytologic smears was HPV 16, followed by HPV 58 and 66. Anti-VLPs was positive in 19.7% and the frequent anti-VLPs type was against HPV 18, followed by HPV 31 and 16. The concordance between the markers for each specific HPV type was noted in 10 women and HPV 16 was the most frequent one. The incidence of multiple HPV infection was 18.9% and that of multiple anti-VLPs antibodies was 31%. Among 103 self-reported virgins, 4.9% had anti-VLP antibodies.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.587-601
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• 2013

During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2795-2798
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• 2012

Objective: This study is conducted to evaluate the effects of anti-HER-2${\times}$anti-CD3 bi-specific antibodies(BsAb) on HER-2/neuover-expressing human colorectal carcinoma cells. Methods: Growth was assessed by MTT assays after exposure of HCT-116 cells to Herceptin, anti-CD3 and BsAb antibodies. Immunocytochemistry was applied to test the HER-2 level of HCT-116. In a nude mouse model, HER-2${\times}$CD3 BsAb was combined with effector cells (peripheral blood lymph cells from normal human being) for observations on in Vivo growth of tumors. Results: Compared with the control group, using effector cells combined with anti-CD3 McAb, Herceptin or HER2${\times}$CD3 BsAb, tumor cell growth in vitro and in vivo was significantly inhibited (P<0.05), most remarkably in the HER2${\times}$CD3 BsAb case. The growth of xenografts with HER2${\times}$CD3 BsAb combined with effector cells was also significantly inhibited when compared with the anti-CD3 McAb or Herceptin groups (P<0.05). Conclusion: HER-2/neu might be a useful target for immunotherapy in colorectal carcinoma, anti-HER2${\times}$anti-CD3 BsAb exerting clear anti-tumor effects.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.905-910
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• 2016

Breast cancer is one of the major threats to female health, and its incidence is rapidly increasing in many countries. Currently, breast cancer is treated with surgery, followed by chemotherapy or radiation therapy, or both. However, a substantial proportion of breast cancer patients might have a risk for local relapse that leads to recurrence of their disease and/or metastatic breast cancer. Therefore searching for new and potential strategies for breast cancer treatment remains necessary. Immunotherapy is an attractive and promising approach that can exploit the ability of the immune system to identify and destroy tumors and thus prevent recurrence and metastatic lesions. The most promising and attractive approach of immunotherapeutic research in cancer is the blockade of immune checkpoints. In this review, we discuss the potential of certain inhibitors of immune checkpoints, such as antibodies targeting cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed death 1 (PD-1) and lymphocyte activation gene-3 (LAG-3), in breast cancer therapeutics. Immune checkpoint inhibitors may represent future standards of care for breast cancer as monotherapy or combined with standard therapies.

• Toxicological Research
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• 제12권1호
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• pp.81-86
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• 1996

Antigenicity of recombinant human interferon $\beta$(LB00013), a newly developed drug for anti-cancer and anti-viral therapeutic use, was investigated in mice and guinea pigs. The following results were obtained: 1. Mice showed no production of antibodies against LB00013 sensitized with aluminum hydroxide gel (Alum) as an adjuvant, when judged by the heterologous passive cutaneous anaphylaxis (PCA) test in rats. Meanwhile, antibodies against ovalbumin (OVA) sensitized with Alum were clearly detected. 2. In guinea pigs, the sensitization of neither LB00013 only nor LB00013 with complete Freund's adjuvant (CFA) produced positive reactions in the homologous active systemic anaphylaxis (ASA) and the PCA tests. Meanwhile, the sensitization of OVA with CFA produced positive reactions in both PCA and ASA. 3. A LB00013-specific reaction was not observed in an indirect hemagglutination(IHA) assay using sera isolated from LB00013 sensitized mice. The present results suggested that LB00013 may have no antigenic potential in mice and guinea pigs.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.483-489
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• 2002

The role of $G{\alpha}12\;and\;G{\alpha}13$ in modulating the IgE receptor-mediated histamine secretion in the streptolysin-o-permeabilized human cultured mast cell was investigated. The expression of $G{\alpha}12\;and\;G{\alpha}13$ proteins were regulated during human cultured mast cell differentiation, and a significant correlation was observed between the levels of expression of $G{\alpha}12\;and\;G{\alpha}13$ proteins and IgE receptor-mediated histamine secretion capability in human cultured mast cells. Antibodies against $G{\alpha}12\;and\;G{\alpha}13$ effectively inhibited the IgE receptor-induced histamine release, and the concentration of anti-$G{\alpha}12$ antibody used to inhibit histamine secretion was shown to also inhibit the IgE receptor-mediated elevation of intracellular $Ca^2+$. Therefore, the results suggest that $G{\alpha}12\;and\;G{\alpha}13$ play roles in modulating IgE receptor-activated $Ca^2+$ influx, thereby regulating histamine release in cultured human mast cells. This is the first report to show that $G{\alpha}12\;and\;G{\alpha}13$ are involved in the regulation of $Ca^2+$ mediated exocytosis in human cultured mast cells.

• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.71-80
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• 1990

항정자항체를 검사할 수 있는 면역분석법개발을 위하여 immunoaffinity chromatography로 분리 정제한 정자표현항원을 microtiter plate에 고정화 시킨것과 효소와 화학발광체로 표지된 2차항체를 사용하여 ELISA법과 CIA법을 개발하고 이 방법의 이용 가능성을 검토하기 위하여 기존 방법인 Kibrick test법으로 임상소견이 다른 남성혈청을 분석 비교하여 다음과 같은 결과를 얻었다. 1. 인간정자 표면항원을 분리하기 위한 immunoaffinity column을 제작키 위하여 인간정자를 토끼에 주사하여 정자에 대한 항혈청을 생산하였으며 이를 Protein A-Sepharose column으로 분리 정제하여 CNBr activated Sepharose-4B에 coupling시켜 immunoaffinity column을 제작하였다. 이 column에 균질화된 정자를 반응시키고 SDS를 넣은 Tris-HCI buffer로 용출시켰을때 60KD정도의 분자량을 갖는 분획을 얻었다. 2. 분리 정제된 human IgG를 microtier plate에 농도를 달리하여 고정화 시키고 ELISA용 Goat anti-human IgG-HRP conjugate와 CIA용 Rabbit anti-human IgG-ABEI-H conjugate와 반응시켜 그 활성도를 측정하였던 바 농도에 따라 반응의 정도가 감소하였다. 3. ELISA법으로 양성혈청의 희석곡선을 작성하였을때 경사가 완만한 것과 급한 것의 두 종류의 경향을 띈 곡선으로 대별되었으며 완만한 경사를 나타내는 것에서 1:160 희석치에서 O.D가가 0.1이하를 음성, 0.1이상 0.2이하를 약양성(weak positive) 그리고 0.2이상을 양성으로 판별하였다. 4. ELISA, CIA 그리고 Kibrick test로 동일시료를 분석 비교하였던바 ELISA와 CIA는 거의 동일한 상관관계를 보였으나 Kibrick test와는 50% 수준만 일치함을 보였다.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.17-24
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• 1998

We have previously demonstrated that specific antigens involved in autoimmunity in endometriosis may be endometrial proteins with molecular weight (mw) of 71, 92, and 103 kilodalton (kDa). The purposes of this study were to determine the incidence of IgG antibodies against these endometrial antigens in peritoneal fluid of patients with endometriosis and to evaluate the antigenic differences between the endometria of patients with and without endometriosis. Forty peritoneal fluid (PF) from 24 patients with endometriosis and 16 patients without endometriosis (control patients) were tested against endometrial protein from patients (n=8) with endometriosis and from control patients (n=10) by western blot. Fifteen (62.5%) of 24 PF samples from patients with endometriosis had specific Immunoglobuiin (Ig) G antibodies against one of three endometrial proteins with mw of 71, 92 and 103 kDa but none of PF samples from control patients had these antibodies. The electrophoretic pattern of endometrial proteins from patients with endometriosis was similiar to that from control patients. Furthemore there was no significant difference in specific PF Immunoglobulin G binding to endometrial proteins regardless of origin of these proteins. Our data indicate that specific humoral immune response can be found in PF of patients with endometriosis and that specific antigens inducing this immune response are present in human endometrium and that there is no antigenic difference between the endometria of patients with and without endometriosis.

• IMMUNE NETWORK
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• 제4권1호
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• pp.16-22
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• 2004

Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.