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791.
Julie A Douthwaite Sudharsan Sridharan Catherine Huntington Jayne Hammersley Rose Marwood Jonna K Hakulinen Margareta Ek Tove Sj?gren David Rider Cyril Privezentzev Jonathan C Seaman Peter Cariuk Vikki Knights Joyce Young Trevor Wilkinson Matthew Sleeman Donna K Finch David C Lowe Tristan J Vaughan 《MABS-AUSTIN》2015,7(1):152-166
Therapeutic monoclonal antibodies targeting G-protein-coupled receptors (GPCRs) are desirable for intervention in a wide range of disease processes. The discovery of such antibodies is challenging due to a lack of stability of many GPCRs as purified proteins. We describe here the generation of Fpro0165, a human anti-formyl peptide receptor 1 (FPR1) antibody generated by variable domain engineering of an antibody derived by immunization of transgenic mice expressing human variable region genes. Antibody isolation and subsequent engineering of affinity, potency and species cross-reactivity using phage display were achieved using FPR1 expressed on HEK cells for immunization and selection, along with calcium release cellular assays for antibody screening. Fpro0165 shows full neutralization of formyl peptide-mediated activation of primary human neutrophils. A crystal structure of the Fpro0165 Fab shows a long, protruding VH CDR3 of 24 amino acids and in silico docking with a homology model of FPR1 suggests that this long VH CDR3 is critical to the predicted binding mode of the antibody. Antibody mutation studies identify the apex of the long VH CDR3 as key to mediating the species cross-reactivity profile of the antibody. This study illustrates an approach for antibody discovery and affinity engineering to typically intractable membrane proteins such as GPCRs. 相似文献
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Neutralizing Activity of Broadly Neutralizing Anti-HIV-1 Antibodies against Primary African Isolates
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C. S. Buer M. J. Correll T. C. Smith M. J. Towler P. J. Weathers M. Nadler J. Seaman D. Walcerz 《In vitro cellular & developmental biology. Plant》1996,32(4):299-304
Summary A nutrient-mist bioreactor was designed that separates the nutrient medium from the electronic components via an acoustic
window. This eliminates compromising culture sterility when repairing mechanical failures common with commercially available
mist reactors. The experimental mist bioreactor is low cost and can be assembled in any laboratory. Toxicity tests of several
potential acoustically transparent materials are included. Details of the construction procedures include methods for casting
the window. Growth data using the newly designed nutrient mist bioreactor are compared to data from a commercial mist reactor,
shake flasks, and Gelrite cultures.Artemisia annua hairy roots andNephrolepis exaltata shoot cultures showed growth comparable to the conventional tissue culture methods. 相似文献
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Identification and characterization of a cell-surface molecule that is selectively induced on rat lymphokine-activated killer cells 总被引:3,自引:0,他引:3
J B Imboden E C Eriksson M McCutcheon C W Reynolds W E Seaman 《Journal of immunology (Baltimore, Md. : 1950)》1989,143(9):3100-3103
We have identified a 40- to 45-kDa cell-surface molecule designated gp42, that is expressed in high levels by rat lymphokine-activated killer (LAK) cells of NK cell origin. gp42 cannot be detected on the precursors of LAK cells and is not present on resting or activated T cells. Rather, expression of gp42 is selectively induced on NK cells by the high concentrations of rIL-2 that are required for the induction of LAK activity. Although the function of gp42 is not known, the selective nature of its expression suggests a role for this molecule in regulating responses that are unique to IL-2-activated NK cells. 相似文献