Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies.

The O-antigen polysaccharide (O-PS) component of lipopolysaccharides on the surface of gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies elicited by O-PS often confer protection against infection; therefore, O-PS glycoconjugate vaccines have proven useful against a number of different pathogenic bacteria. However, conventional methods for natural extraction or chemical synthesis of O-PS are technically demanding, inefficient, and expensive.

Development of purification processes for fully human bispecific antibodies based upon modification of protein A binding avidity.

There is strong interest in the design of bispecific monoclonal antibodies (bsAbs) that can simultaneously bind 2 distinct targets or epitopes to achieve novel mechanisms of action and efficacy. Multiple bispecific formats have been proposed and are currently under development. Regeneron's bispecific technology is based upon a standard fully human IgG antibody in order to minimize immunogenicity and improve the pharmacokinetic profile.

Production of secretory and extracellular N-linked glycoproteins in Escherichia coli.

The Campylobacter jejuni pgl gene cluster encodes a complete N-linked protein glycosylation pathway that can be functionally transferred into Escherichia coli. In this system, we analyzed the interplay between N-linked glycosylation, membrane translocation and folding of acceptor proteins in bacteria. We developed a recombinant N-glycan acceptor peptide tag that permits N-linked glycosylation of diverse recombinant proteins expressed in the periplasm of glycosylation-competent E.

Infrared hole burning of crown ether 18-c-6 ammonium ion complexes.

Crystals of the complexes of 18-crown-6 with a number of ammonium salts (bromide, chloride, two forms of iodide, nitrate) and with amminetrifluoroboron have been prepared, analyzed with X-ray diffraction, and then investigated by infrared hole burning. The complexes all have similar hydrogen-bonding arrangements between the crown ether ring and the ammonium nitrogen. Hole burning of the infrared bands at low temperature identifies the N-D hydrogen bonds of the compounds that have been doped with a small amount of deuterium.

Infrared hole burning and conformational change in a borane-ammonia complex.

The N-D stretching region in the infrared spectrum of the ammonia complex of tris-(2-methoxymethyl-phenol)-borane containing one D atom has been examined. The N-D bands have been hole burned, and the resulting spectra reveal the reorientation kinetics of the ammonia. The ammonia is hydrogen bonded with the bond distances and reorientation barrier typical of other compounds.