• Title/Summary/Keyword: human antibodies

Search Result 449, Processing Time 0.029 seconds

Resurrection of antibody as a therapeutic drug (항체 : 치료제로서의 부활)

  • Chung, Hong Keun;Chung, Junho
    • IMMUNE NETWORK
    • /
    • v.1 no.1
    • /
    • 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.

  • PDF

Cross-reactivity of Human Polyclonal Anti-GLUT1 Antisera with the Endogenous Insect Cell Glucose Transporters and the Baculovirus-expressed GLUT1

  • Lee, Chong-Kee
    • Biomedical Science Letters
    • /
    • v.7 no.4
    • /
    • pp.161-166
    • /
    • 2001
  • Most mammalian cells take up glucose by passive transport proteins in the plasma membranes. The best known of these proteins is the human erythrocyte glucose transporter, GLUT1. High levels of heterologous expression far the transporter are necessary for the investigation of its three-dimensional structure by crystallization. To achieve this, the baculovirus expression system has become popular choice. However, Spodoptera frugiperda Clone 9 (Sf9) cells, which are commonly employed as the host permissive cell line to support baculovirus replication and protein synthesis, grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, suggesting the presence of endogenous glucose transporters. Furthermore, very little is known of the endogenous transporters properties of Sf9 cells. Therefore, human GLUT1 antibodies would play an important role for characterization of the GLUT1 expressed in insect cell. However, the successful use of such antibodies for characterization of GLUT1 expression m insect cells relies upon their specificity for the human protein and lack of cross-reaction with endogenous transporters. It is therefore important to determine the potential cross-reactivity of the antibodies with the endogenous insect cell glucose transporters. In the present study, the potential cross-reactivity of the human GLUT1 antibodies with the endogenous insect cell glucose transporters was examined by Western blotting. Neither the antibodies against intact GLUT1 nor those against the C-terminus labelled any band migrating in the region expected fur a protein of M$_r$ comparable to GLUT1, whereas these antibodies specifically recognized the human GLUT1. Specificity of the human GLUT1 antibodies tested was also shown by cross-reaction with the GLUT1 expressed in insect cells. In addition, the insect cell glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

  • PDF

Antibody Engineering

  • Hong, Hyo-Jeong;Kim, Sun-Taek
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.7 no.3
    • /
    • pp.150-154
    • /
    • 2002
  • Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. An indication of the emerging significance of antibody-based therapeutics is that over a third of the proteins currently undergoing clinical trials in the United States are antibodies. Until the late 1980's, antibody technology relied primarily on animal immunization and the expression of engineered antibodies. However, the development of methods for the expression of antibody fragments in bacteria and powerful techniques for screening combinatorial libraries, together with the accumulating structure-function data base of antibodies, have opened unlimited opportunities for the engineering of antibodies with tailor-made properties for specific applications. Antibodies of low immunogenicity, suitable for human therapy and in vivo diagnosis, can now be developed with relative ease. Here, antibody structure-function and antibody engineering technologies are described.

Preparation of Dopamine Transporter-specific Antibodies Using Molecular Cloned Genes

  • Lee, Shee-Yong;Im, Suhn-Young;Kim, Kyeong-Man
    • Archives of Pharmacal Research
    • /
    • v.22 no.3
    • /
    • pp.262-266
    • /
    • 1999
  • Dopamine transporter (DAT) plays the most important role in terminating the actions of dopamines released into the synaptic cleft. DAT is also the target of various psychotropic drugs such as cocaine and amphetamine. In this study were prepared DAT-specific antibodies using the 2nd extracellular loop of rat DAT as an antigen. The 2nd extracellular loop of the rat DAT was expressed in bacterial as a fusion protein with glutathione-S-transferase, and injected ito rabbits to raise antibodies. Produced antibodies clearly recognized the rat DAT in ELISA, immunoblotting, and immumoprecipitation. As expected from the high sequence homology between the rat and human DAT, the antibodies raised for the rat DAT cross-reacted with the human DAT in the immunoblotting. Considering the specificity for DAT with wide range of applications such as ELISA, immunoblotting, and immunoprecipitation, these antibodies would be valuable tool for understanding the pharmacological actions of dopamine transporter and drug addition.

  • PDF

Immunogenecity of Low Molecular Weight Immunogens in Laying Hens (닭에서 저급 분자량 항원의 면역원성)

  • Lee, Kyong-Ae
    • Korean Journal of Human Ecology
    • /
    • v.5 no.1
    • /
    • pp.75-80
    • /
    • 1996
  • The immunogenecity of low molecular weight ($MW{\le}30,000$) immunogens in laying hens was investigated. Immunogens were insulin derivatives and ${\beta}$-lactoglobulin(${\beta}-Lg$). Insulin derivatives were reduced- carboxymethylated(RCM) insulin, and RCM-A and RCM-B chain of insulin. The yolk antibodies against RCM-A chain of insulin appeared after the first immunization. The yolk antibodies against RCM-B chain of insulin were elicited 5 weeks after the third booster injection. Although the anti-RCM-B chain yolk antibodies recognized native insulin, the anti-RCM-A chain yolk antibodies didn't native insulin. The anti-RCM insulin yolk antibodies were induced after the second booster injection and showed cross-reactivities with native insulin. On the other hand, ${\beta}-Lg$ showed stronger immunogenecity than insulin derivatives. The $anti-{\beta}-Lg$ yolk antibodies were produced after the second booster injection and the peak titer was reached 3 weeks after the third booster injection.

  • PDF

Flow cytometric immunophenotyping of canine adipose-derived mesenchymal stem cells (ADMSCs) and feline ADMSCs using anti-human antibodies

  • Ko, Minho;Lee, Kwon Young;Kim, Sae Hoon;Kim, Manho;Choi, Jung Hoon;Im, Wooseok;Chung, Jin Young
    • Korean Journal of Veterinary Research
    • /
    • v.58 no.1
    • /
    • pp.33-37
    • /
    • 2018
  • Various trials have been conducted to develop therapies for serious untreatable diseases. Among these, those using stem cells have shown great promise, and adipose-derived mesenchymal stem cells (ADMSCs) are easier to obtain than other types of stem cells. Prior to clinical trials, characterization of ADMSCs with monoclonal antibodies should be performed. However, it is difficult to use species-specific antibodies for veterinarians. This study was conducted to confirm the panel of human antibodies applicable for use in immunophenotypic characterization of canine adipose-derived stem cells and feline ADMSCs extracted from subcutaneous adipose tissue collected during ovariohysterectomy. For flow cytometric immunophenotyping, the third passages of canine ADMSC and feline ADMSC and human CD31, CD34, CD42, CD44, CD62 and CD133 antibodies were used. Of these, CD133 reacted with canine cells (3.74%) and feline cells (1.34%). CD133 is known as a marker related with more primitive stem cell phenotype than other CD series. Because this human CD133 was not a species-specific antibody, accurate percentages of immunoreactivity were not confirmed. Nevertheless, the results of this study confirmed human CD133 as a meaningful marker in canine and feline ADMSCs.

Effects of Monoclonal Antibodies against Human Stathmin Combined with Paclitaxel on Proliferation of the QG-56 Human Lung Carcinoma Cell Line

  • Yuan, Shao-Fei;Chen, Wen-Jun;Zhu, Lin-Jia;Zheng, Wei-E.;Chen, Hua;Xiong, Jian-Ping
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.6
    • /
    • pp.2967-2971
    • /
    • 2012
  • Objective: To explore whether monoclonal antibodies against stathmin and the chemotherapuetic agent paclitaxel have synergenic effects in inhibiting growth and inducing apoptosis in human QG-56 cells. Methods: QG-56 cells were treated with monoclonal antibodies against stathmin or paclitaxel alone or in combination, with untreated cells used as controls. After 24, 48, 72 and 96 hours the cell growth condition was observed under an inverted microscope and inhibition was studied by MTT assay; apoptosis was analyzed by flow cytometry. Results: The populations decreased and cell shape and size changed after the various treatments. Monoclonal antibodies against stathmin and paclitaxel used alone or incombination inhibited the proliferation of QG-56 cells, especially in combination with synergism (P<0.05). Combined treatment also resulted in a significantly higher apoptosis rate than in the other groups (P<0.05). Conclusions: Monoclonal antibodies against stathmin and paclitaxel used alone or in combination can inhibit proliferation of QG-56 cells and induce apoptosis when applied together. The observed synergistic effects may have important implications for clinical application.

Human Cardiac Troponin I 면역분석을 위한 단일클론 항체의 특성화

  • O, Heung-Il;Yang, Jin-A;Baek, Ui-Hwan;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
    • /
    • 2000.11a
    • /
    • pp.713-714
    • /
    • 2000
  • Six monoclonal antibodies to human cardiac troponin I (hcTnI) were produced to eventually develop an immunosensor for acute myocardial infarction (AMI). For the characterization of these antibodies, a set of 11 different peptides covering selected ranges of the complete amino acid sequence of hcTnI was prepared and used for epitope mapping. Such analysis allowed to select an appropriate pair of antibodies that can form a sandwich type of immune complexes and was consequently used for an immunoassay.

  • PDF

Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
    • /
    • v.32 no.5
    • /
    • pp.474-479
    • /
    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

  • PDF

Development of a Quantitative ELISA for Anti HER-2 Antibodies using Human HER-2 Recombinant Proteins (인간 HER-2 재조합 단백질을 사용한 항 HER-2 항체 단백질의 ELISA 정량 방법 개발)

  • Jung, Sun-Ki;Ryu, Chang-Seon;Choung, Kyu-Jin;Song, Gyu-Yong;Kim, Sang-Kyum
    • YAKHAK HOEJI
    • /
    • v.55 no.1
    • /
    • pp.16-21
    • /
    • 2011
  • HER-2 (Human Epidermal Growth Factor Receptor-2) is a protein giving higher aggressiveness in human breast cancers. Trastuzumab is a monoclonal antibody that targets HER-2 and is known to extend survival across all stages of HER2-positive breast cancer. In this study, we attempted to development of a quantitative ELISA (Enzyme-Linked ImmunoSorbent Assay) for evaluating anti HER-2 antibodies using human HER-2 recombinant proteins to support antibody producing processes and pharmacokinetic studies. We established direct or indirect ELISA method for the trastuzumab-like protein combined human recombinant HER-2. The ELISA method will prove to be great value in quantitating anti-HER-2 antibodies levels for developing anticancer antibodies.