'A nonimmune library of 10(9) human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow-cytometric sorting. The yeast library can be amplified 10(10)-fold without measurable loss of clonal diversity, allowing its effectively indefinite expansion. The ... More
'Array-based sensors provide an architecture for multianalyte sensing. In this paper, we report a new approach for array fabrication. Sensors are made by immobilizing different reactive chemistries on the surfaces of microspheres. Sensor arrays are prepared by randomly distributing a mixture of microsphere sensors on an optical substrate containing thousands ... More
Interphase cytogenetics in pathology: principles, methods, and applications of fluorescence in situ hybridization (FISH).
AuthorsWerner M, Wilkens L, Aubele M, Nolte M, Zitzelsberger H, Komminoth P
JournalHistochem Cell Biol
PubMed ID9387931
'Characteristic chromosome aberrations have been identified in various tumors. Fluorescence in situ hybridization (FISH) using specific probes that are generated by vector cloning or in vitro amplification and labeled with fluorescent dyes allow for the detection of these genetic changes in interphase cells. This technique, that is also referred to ... More
Improved immunocytochemical detection of biotinylated probes with Neutralite avidin.
Authorsvan Roy N, Mangelschots K, Speleman F
JournalTrends Genet
PubMed ID8488564
Adhesive and mammalian transglutaminase substrate properties of Candida albicans Hwp1.
AuthorsStaab JF, Bradway SD, Fidel PL, Sundstrom P
JournalScience
PubMed ID10066176
The pathogenesis of candidiasis involves invasion of host tissues by filamentous forms of the opportunistic yeast Candida albicans. Morphology-specific gene products may confer proinvasive properties. A hypha-specific surface protein, Hwp1, with similarities to mammalian small proline-rich proteins was shown to serve as a substrate for mammalian transglutaminases. Candida albicans strains ... More
Biomolecular interactions at phospholipid-decorated surfaces of liquid crystals.
AuthorsBrake JM, Daschner MK, Luk YY, Abbott NL
JournalScience
PubMed ID14684814
The spontaneous assembly of phospholipids at planar interfaces between thermotropic liquid crystals and aqueous phases gives rise to patterned orientations of the liquid crystals that reflect the spatial and temporal organization of the phospholipids. Strong and weak specific-binding events involving proteins at these interfaces drive the reorganization of the phospholipids ... More
Engineering chemical reactivity on cell surfaces through oligosaccharide biosynthesis.
AuthorsMahal LK, Yarema KJ, Bertozzi CR
JournalScience
PubMed ID9173543
Cell surface oligosaccharides can be engineered to display unusual functional groups for the selective chemical remodeling of cell surfaces. An unnatural derivative of N-acetyl-mannosamine, which has a ketone group, was converted to the corresponding sialic acid and incorporated into cell surface oligosaccharides metabolically, resulting in the cell surface display of ... More
Coupling of biotin-(poly(ethylene glycol))amine to poly(D,L-lactide-co-glycolide) nanoparticles for versatile surface modification.
AuthorsWeiss B, Schneider M, Muys L, Taetz S, Neumann D, Schaefer UF, Lehr CM
JournalBioconjug Chem
PubMed ID17590034
Generally, polymeric nanoparticles (NP) for drug targeting are designed to entrap the drug moiety in the core and to present the targeting moiety on the surface. However, in most cases, common preparation techniques of polymeric NP need to be specifically arranged for each compound to be entrapped or attached. In ... More
Linear V1-vascular vasopressin antagonists suitable for radioiodination, biotinylation, and fluorescent labeling.
AuthorsThibonnier M, Bayer AL, Madhun Z
JournalAm J Physiol
PubMed ID8279546
We modified several linear V1-vascular arginine vasopressin (AVP) antagonists to obtain compounds suitable for radioiodination, biotinylation, and fluorescent labeling. In binding competition experiments with human platelet V1-vascular AVP receptors, the linear V1 antagonist phenylacetyl-D-Tyr(Et)-Phe-Gln-Asn-Lys-Pro-Arg-NH2 (PhaaGln) displayed the greatest affinity [dissociation constant (Kd) = 0.05 +/- 0.01 nM]. The radioiodinated compound ... More
Novel biotinylated plasmid expression vectors retain biological function and can bind streptavidin.
AuthorsLeahy P, Carmichael GG, Rossomando EF
JournalBioconjug Chem
PubMed ID8889015
A new method for coupling proteins to plasmid expression vectors is presented. Biotin was covalently attached to a plasmid expression vector containing a chloramphenicol acetyltransferase (CAT) gene. The specific label was one biotin per 100 bp. An electrophoretic mobility shift assay showed that the plasmid was capable of binding multiple ... More
Preservation of NADH voltammetry for enzyme-modified electrodes based on dehydrogenase.
AuthorsHayes MA, Kuhr WG
JournalAnal Chem
PubMed ID10330904
Minimizing overpotential and generating high faradaic currents are critical issues for fast-scan voltammetry of beta-nicotinamide adenine dinucleotide (NADH) for the sensitivity of enzyme-modified electrodes based on dehydrogenases. Although NADH voltammetry exhibits high overpotential and poor voltammetric peak shape at solid electrode surfaces, modification of the electrode surface can improve the ... More
Tracking movements of lipids and Thy1 molecules in the plasmalemma of living fibroblasts by fluorescence video microscopy with nanometer scale precision.
AuthorsHicks BW, Angelides KJ
JournalJ Membr Biol
PubMed ID7658460
The lateral diffusion of 100 nm fluorescent latex microspheres (FS) bound to either N-biotinyl-phosphatidyl-ethanolamine or the glycosylphosphatidylinositol-linked protein Thy1 were monitored in the plasmalemma of primary rat fibroblasts by single particle tracking of FS centroids from digital fluorescence micrographs. A silicon intensified target camera was found to be superior to ... More
Molecular cloning, sequencing, and functional expression of a cDNA encoding the human V1a vasopressin receptor.
AuthorsThibonnier M, Auzan C, Madhun Z, Wilkins P, Berti-Mattera L, Clauser E
JournalJ Biol Chem
PubMed ID8106369
Vasopressin (AVP), the antidiuretic hormone, is a cyclic nonapeptide that acts through binding to G protein-coupled specific membrane receptors pharmacologically divided into three subtypes (V1a, V1b, and V2) linked to distinct second messengers. Within the family of human AVP receptors, the V2 AVP receptor has been cloned, but the structure ... More