TY - JOUR A1 - Holzlöhner, Pamela A1 - Hanack, Katja T1 - Generation of murine monoclonal antibodies by hybridoma technology JF - JoVE : Video journal N2 - Monoclonal antibodies are universal binding molecules and are widely used in biomedicine and research. Nevertheless, the generation of these binding molecules is time-consuming and laborious due to the complicated handling and lack of alternatives. The aim of this protocol is to provide one standard method for the generation of monoclonal antibodies using hybridoma technology. This technology combines two steps. Step 1 is an appropriate immunization of the animal and step 2 is the fusion of B lymphocytes with immortal myeloma cells in order to generate hybrids possessing both parental functions, such as the production of antibody molecules and immortality. The generated hybridoma cells were then recloned and diluted to obtain stable monoclonal cell cultures secreting the desired monoclonal antibody in the culture supernatant. The supernatants were tested in enzyme-linked immunosorbent assays (ELISA) for antigen specificity. After the selection of appropriate cell clones, the cells were transferred to mass cultivation in order to produce the desired antibody molecule in large amounts. The purification of the antibodies is routinely performed by affinity chromatography. After purification, the antibody molecule can be characterized and validated for the final test application. The whole process takes 8 to 12 months of development, and there is a high risk that the antibody will not work in the desired test system. KW - Immunology KW - Issue 119 KW - monoclonal antibodies KW - hybridoma technology KW - myeloma cells KW - B lymphocytes KW - antigen KW - immunconjugate Y1 - 2017 U6 - https://doi.org/10.3791/54832 SN - 1940-087X IS - 119 PB - JoVE CY - Cambridge ER - TY - JOUR A1 - Lütkecosmann, Steffi A1 - Warsinke, Axel A1 - Tschöpe, Winfried A1 - Eichler, Rüdiger A1 - Hanack, Katja T1 - A novel monoclonal antibody suitable for the detection of leukotriene B4 JF - Biochemical and biophysical research communications N2 - Leukotriene B4 as an inflammatory mediator is an important biomarker for different respiratory diseases like asthma, chronic obstructive pulmonary disease or cystic lung fibrosis. Therefore the detection of LTB4 is helpful in the diagnosis of these pulmonary diseases. However, until now its determination in exhaled breath condensates suffers from problems of accuracy. Reasons for that could be improper sample collection and preparation methods of condensates and the lack of consistently assay specificity and reproducibility of the used immunoassay detection system. In this study we describe the development and the characterization of a specific monoclonal antibody (S27BC6) against LTB4, its use as molecular recognition element for the development of an enzyme-linked immunoassay to detect LTB4 and discuss possible future diagnostic applications. KW - Leukotriene B4 KW - Monoclonal antibody KW - Immunosensor KW - Chronic obstructive pulmonary disease (COPD) KW - Hapten Y1 - 2017 U6 - https://doi.org/10.1016/j.bbrc.2016.11.157 SN - 0006-291X SN - 1090-2104 VL - 482 IS - 4 SP - 1054 EP - 1059 PB - Elsevier CY - San Diego ER - TY - JOUR A1 - Roggenbuck, Dirk A1 - Goihl, Alexander A1 - Hanack, Katja A1 - Holzloehner, Pamela A1 - Hentschel, Christian A1 - Veiczi, Miklos A1 - Schierack, Peter A1 - Reinhold, Dirk A1 - Schulz, Hans-Ulrich T1 - Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2 JF - Clinical chemistry and laboratory medicine : journal of the Forum of the European Societies of Clinical Chemistry - the European Branch of the International Federation of Clinical Chemistry and Laboratory Medicine N2 - To better understand emerging adults’ perceptions of family interactions and value transmission to the next generation, we examined Hmong American emerging adults’ reflections on their parents’ parenting. Participants discussed what parenting practices they would do differently and others they hoped to emulate with their future adolescent children. Thirty Hmong American emerging adults (18-25 years; M = 21.2 years; 50% female) participated in interviews that focused retrospectively on the parent–adolescent relationship. Results revealed that emerging adults wanted to parent differently in three ways: less pressure about education, fewer restrictions, and more open communication. Emerging adults imagined being a similar parent in four ways: promoting education, promoting life values, giving guidance, and offering love and support. The findings highlight parenting practices that Hmong American emerging adults plan on transmitting (and not transmitting) to their own children, offering a glimpse into the type of parents the emerging adults may become. KW - acute pancreatitis KW - chronic pancreatitis KW - GP2 isoform alpha KW - pancreatic neoplasms KW - severe acute pancreatitis KW - zymogen granule membrane glycoprotein GP2 Y1 - 2017 U6 - https://doi.org/10.1515/cclm-2016-0797 SN - 1434-6621 SN - 1437-4331 VL - 55 SP - 854 EP - 864 PB - De Gruyter CY - Berlin ER -