muramidase and glycolic-acid

Nanoparticles of cadmium telluride (CdTe) coated with thioglycolic acid (TGA) were prepared in the water phase. The interaction between CdTe nanoparticles (NPs) and lysozyme (Lyz) was investigated by fluorescence and circular dichroism (CD) spectroscopy at pH 7.40. It was proved that the fluorescence quenching of Lyz by CdTe NPs was mainly a result of the formation of CdTe-Lyz complex. By the fluorescence quenching results, the Stern-Volmer quenching constant (K(SV)), binding constant (Ka) and binding sites (n) were calculated. The binding distance (r) between Lyz (the donor) and CdTe NPs (the acceptor) was obtained according to fluorescence resonance energy transfer (FRET). Gradual addition of CdTe NPs to the solution of Lyz led to a marked increase in fluorescence polarization (P) of Lyz, which indicated that CdTe NPs were located in a restricted environment of Lyz. The effect of CdTe NPs on the conformation of Lyz has been analyzed by means of synchronous fluorescence spectra and CD spectra, which provided the evidence that the secondary structure of Lyz has been changed by the interaction of CdTe NPs with Lyz.

Topics: Binding Sites; Cadmium Compounds; Circular Dichroism; Glycolates; Kinetics; Models, Statistical; Muramidase; Nanoparticles; Nanotechnology; Protein Binding; Protein Conformation; Protein Structure, Secondary; Spectrometry, Fluorescence; Spectrophotometry; Tellurium; Water

When encapsulating proteins in polymer microspheres for sustained drug delivery there are three stages during which the stability of the protein must be maintained: (1) the fabrication of the microspheres, (2) the storage of the microspheres, and (3) the release of the encapsulated protein. This study focuses on the effects of polymer degradation products on the primary, secondary, and tertiary structure of tetanus toxoid, ovalbumin (Ova), and lysozyme after incubation for 0 or 20 days in the presence of ester (lactic acid and glycolic acid) and anhydride (sebacic acid and 1,6-bis(p-carboxyphenoxy)hexane) monomers. The structure and antigenicity or enzymatic activity of each protein in the presence of each monomer was quantified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism, and fluorescence spectroscopy were used to assess/evaluate the primary, secondary, and tertiary structures of the proteins, respectively. Enzyme-linked immunosorbent assay was used to measure changes in the antigenicity of tetanus toxoid and Ova and a fluorescence-based assay was used to determine the enzymatic activity of lysozyme. Tetanus toxoid was found to be the most stable in the presence of anhydride monomers, while Ova was most stable in the presence of sebacic acid, and lysozyme was stable when incubated with all of the monomers studied.

Topics: Animals; Biocompatible Materials; Cattle; Chemistry, Pharmaceutical; Chickens; Circular Dichroism; Decanoic Acids; Dicarboxylic Acids; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Glycolates; Hexanes; In Vitro Techniques; Lactic Acid; Microspheres; Models, Statistical; Muramidase; Polyanhydrides; Polymers; Proteins; Spectrometry, Fluorescence; Tetanus Toxin; Time Factors