two sialic acid specific lectins episesarma tetragonum agglutinin–1 and 2 were purified from the hemolymph of the mangrove crab, episesarma tetragonum . the major lectin was purified using cnbr-activated sepharose 4b conjugated to fetuin. n-acetyl glucosamine containing buffer was used for elution. the hemagglutination activity of purified lectin was inhibited by glycoproteins containing siaα, ...

We have developed a novel method for efficient screening of affinity ligands (aptamers) from a complex single-stranded DNA (ssDNA) library by employing single molecule emulsion PCR based on agarose droplet microfluidic technology . Statistically diluted ssDNA of a pre-enriched library was encapsulated into individual uniform agarose droplets for highly efficient single molecule emulsion PCR to ...

Cytochrome c oxidase binds protonophore carbonyl cyanide 3-chlorophenylhydrazone (CCCP) with high affinity. There are 1.46 high-affinity binding sites per cytochrome c oxidase for CCCP with dissociation constant 2.7 x 10(-7) mol/l. The bond between the CCCP and cytochrome c oxidase accomplishes through the group on cytochrome c oxidase with pKa 6.64 and is based on the electrostatic interaction...

responsible for the extended substrate spectrum is the R2 loop (residues 289–310 according to the amino acid numbering system of P99; Figure 1). Another issue that we wish to highlight from our reading of the work of Mammeri et al. is a commonly made oversimplification that a decrease in Km indicates an increased affinity of a substrate for an enzyme. Because an increased k2/k3 ratio (not only ...

We demonstrate that the spatial analysis of nanofluidic-embedded biosensors permits the fast and direct discrimination of single-nucleotide difference (SND) within microRNA sequences in a single step interaction. We first show that the spatial hybridization profile depends on the duplex affinity, and we speculate that the affinity constant can be inferred during target injection, which is not p...

A high-affinity riboflavin -binding protein was isolated and characterized for the first time from pregnant-rat sera by affinity chromatography on a lumiflavin-agarose column. The purified protein was homogeneous by the criteria of analytical polyacrylamide-gel disc electrophoresis, gel-filtration chromatography on Sephadex G-100 and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. I...

Development of protein therapeutics or biosensors often requires in vitro affinity maturation. Here we report a robust affinity engineering strategy using a custom designed library. The strategy consists of two steps beginning with identification of beneficial single amino acid substitutions then combination. A high quality combinatorial library specifically customized to a given binding-interf...

The O2 affinity of fiSH chains is lowered by H+, inositol hexaphosphate (IHP), and COz. As the oxygen affinity of gSH monomers (flISH) is lower than that of fiSH tetramers (@dSH), it is possible that IHP and CO2 exert their influence on the Oz affinity of PSH chains by increasing the dissociation constant of +3dsH rather than by a direct effect on the molecule. In order to test for this hypothe...

1. The equilibrium and kinetics of cyanide binding to ferroperoxidase were investigated. At pH9.1 the equilibrium and kinetic measurements agree closely and disclose a single process with an affinity constant of 1.1x10(3)m(-1) and combination and dissociation velocity constants of 29m(-1).s(-1) and 2.5x10(-2)s(-1) respectively. 2. At pH values below 8 the affinity constant falls until at pH6.0 ...