Search Results - (Author, Cooperation:E. S. Ward)

2012-06-05

Nature Publishing Group (NPG)

0028-0836

1476-4687

Biology

Chemistry and Pharmacology

Medicine

Natural Sciences in General

Physics

Animals ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Disease Models, Animal ; Fetal Blood/cytology/metabolism ; HEK293 Cells ; Hematopoietic Stem Cells/*cytology/*metabolism ; Humans ; Leukemia/*metabolism/*pathology ; Membrane Glycoproteins/genetics/*metabolism ; Mice ; Myeloid-Lymphoid Leukemia Protein ; Receptors, Immunologic/genetics/*metabolism

2018-09-04

Rockefeller University Press

0022-1007

1540-9538

Medicine

Autoimmunity, Metabolism

1365-3083

Blackwell Publishing Journal Backfiles 1879-2005

Medicine

Site-directed mutagenesis of a recombinant Fc-hinge fragment has previously been used to identify a region of the murine IgG 1 molecule that controls catabolism, and this site encompasses amino acid residues at the interface of the CH2 and CH3 domains. In the current study the nature of this ‘catabolic site’ has been further analysed using recombinant techniques. Fc-hinge, CH2-hinge, CH2 and CH3 fragments have been expressed in Escherichia coli, purified and analysed in pharmacokinetic studies in mice. The CH2-hinge has been analysed as both a monomer and dimer, and the dimer has a longer β phase half-life (61.6 h) than the monomer (29.1 h). This suggests that two catabolic sites per Fc fragment are required for serum persistence. The need for two functional sites per molecule has been confirmed by the analysis of a hybrid Fc-hinge fragment comprising a heterodimer of one Fc-hinge with the wild type (WT) IgGl sequence and a mutant Fc-hinge with a defective catabolic site (mutated at His310, Gln311, His433 and Asn434). This hybrid is cleared with a β phase half-life of 37.9 h and this is significantly shorter than that of the WT Fc-hinge fragment (82.9 h). In contrast to the CH2-hinge dimer, the CH3 domain is cleared rapidly (β phase half-life of 21.3 h) indicating that the region of this domain (His433 and Asn434) previously identified as being involved in the control of catabolism is not sufficient in the absence of the CH2 domain for the serum persistence of an IgG fragment. The data extend our earlier observations concerning a region of the murine IgGl molecule that is involved in the control of catabolism and have implications for the design of engineered antibodies for therapy.

Electronic Resource

1365-3083

Blackwell Publishing Journal Backfiles 1879-2005

Medicine

An expression system for the production of recombinant T-cell receptor (TCR) variable domains would, inter alia, allow structural studies lo be carried out and provide protein for the generation of anti-clonotypic antibodies, in this report the Vα and Vβ domain genes have been isolated from a T-cell hybridoma which is associated with the pathogenesis of experimental allergic encephalomyelitis (EAE) In the H-2 mouse. These have been expressed as secreted domains in Escherichia coli, using secretion vectors previously used for the production of immunoglobulin fragments. Both Vα and Vβ domains are secreted in milligram quantities into the culture supernatant, although the levels of the Vα domain are about 10-20 Ibid higher than those of the Vβ domain. This expression system offers a rapid route for the production of recombinant TCRs in soluble form.

Electronic Resource

0017-789X

General, Interdisciplinary

0017-789X

General, Interdisciplinary

0017-789X

General, Interdisciplinary