'Camelising' human antibody fragments: NMR studies on VH domains.
A human heavy chain variable domain (VH) was expressed in bacteria for structural analysis by NMR spectroscopy. NMR analysis was initially impossible due to the short transverse proton relaxation time of the VH, probably caused by aggregation through the exposed interface naturally in contact with the light chain. The relaxation time was improved to normal values when this interface was mutated to mimic heavy chains of camel antibodies naturally devoid of light chains and through the use of the detergent CHAPS. Assignment of NMR signals will now be possible after isotopic labeling. Implications for the design of VH domains as minimum size immunoreagents are outlined.
Davies
J
J
MRC Laboratory of Molecular Biology, Cambridge, UK.
Riechmann
L
L
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002162:Camelus / Q000276:immunology*; D002793:Cholic Acids; D003903:Deuterium; D004355:Drug Stability; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry* / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry; D017385:Sequence Homology
D002793:Cholic Acids; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D003903:Deuterium; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate