[51] Streptavidin-Enzyme complexes in detection of antigens on western blots

Brakel, Christine L.; Brower, Mark S.; Garry, Kimberly E.

doi:10.1016/0076-6879(90)84305-z

Cited by 3 publications

(1 citation statement)

References 13 publications

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“…Streptavidin-coated solid surfaces, on which streptavidin is covalently immobilized, have proven very useful for a variety of biomedical assays, including clinical diagnostics ( − ). The properties of one such streptavidin-coated solid surface () were recently investigated.…”

Section: Resultsmentioning

confidence: 99%

A Streptavidin Mutant Useful for Directed Immobilization on Solid Surfaces

et al. 2001

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A streptavidin mutant has been designed and produced that allows the specific, covalent immobilization of streptavidin on solid surfaces. This streptavidin mutant was constructed by fusing a six-residue sequence, containing a single cysteine, to the carboxyl terminus of streptavidin. Because this mutant has no other cysteine residues, the reactive sulfhydryl group of the cysteine residue serves as a unique immobilization site for conjugation using sulfhydryl chemistry. This streptavidin mutant was efficiently immobilized on maleimide-coated solid surfaces via its unique immobilization site. Characterization of the immobilized streptavidin mutant for the ability to bind to biotinylated macromolecules and the dissociation rates of bound biotin showed that the biotin-binding properties of this mutant were minimally affected by immobilization on solid surfaces. This streptavidin could be readily incorporated into a wide variety of solid-phase diagnostic tests and biomedical assays. This could enhance the performance of streptavidin-based solid-phase assay systems.

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Section: Resultsmentioning

confidence: 99%

A Streptavidin Mutant Useful for Directed Immobilization on Solid Surfaces

et al. 2001

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Detection of Endogenous Biotin-Containing Proteins in Bone and Cartilage Cells with Streptavidin Systems

Praul

Brubaker

Leach

et al. 1998

Biochemical and Biophysical Research Communications

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Increased sialylation and defucosylation of plasma proteins are early events in the acute phase response

2005

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Within hours of turpentine injection to stimulate the acute phase (AP) response in rats, the N-acetylneuraminic acid content of plasma proteins increases and that of fucose decreases, each by about 60%. The two changes are inversely related (r = -0.97). The NeuAc/Gal ratio increases from the normal 0.75 to 1.0 on day 2 of the AP. Whereas 50% of the isolated oligosaccharides of normal plasma proteins are retarded on immobilized Ricinus communis agglutinin, those from day 2 AP plasma fail to do so. This indicates that NeuAc caps the normally Gal-terminated chains. alpha1-Acid glycoprotein (a positive AP protein), alpha1-macroglobulin (a non-AP protein), and alpha1-inhibitor3 (a negative AP protein) also show similar alterations in NeuAc/Gal ratio and decreases in Fuc. alpha2-Macroglobulin, which arises only during the AP, does not contain significant amounts of Fuc. Sambucus nigra agglutinin (alpha2,6-linked NeuAc-specific) binds a majority of plasma proteins, and binding is increased during the AP response. Maackia amurensis lectin (alpha2,3-linked NeuAc-specific) binds only three proteins in normal plasma and three additional proteins in AP plasma. The Fuc-specific Aleuria aurantia agglutinin and Lens culinaris agglutinin each detect five proteins in normal plasma. Their binding decreases during the AP response. These results show that: (1) sialylation and defucosylation of preexisting plasma proteins occur rapidly in the AP response; (2) sialylation caps the preexisting Gal-terminating oligosaccharides; and (3) the oligosaccharides of even the non-AP and negative AP proteins are modified. These changes are distinct from the elevation in the levels of protein-bound monosaccharides and the altered concanavalin A-binding profile the oligosaccharides of AP proteins acquire in diseases.

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[51] Streptavidin-Enzyme complexes in detection of antigens on western blots

Cited by 3 publications

References 13 publications

A Streptavidin Mutant Useful for Directed Immobilization on Solid Surfaces

A Streptavidin Mutant Useful for Directed Immobilization on Solid Surfaces

Detection of Endogenous Biotin-Containing Proteins in Bone and Cartilage Cells with Streptavidin Systems

Increased sialylation and defucosylation of plasma proteins are early events in the acute phase response

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