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1.
Biosens Bioelectron ; 43: 143-7, 2013 May 15.
Article in English | MEDLINE | ID: mdl-23298625

ABSTRACT

A facile route for sensitive label-free detection of bio-toxins using aligned single walled carbon nanotubes is described. This approach involves patterning of a catalyst on the surface of a quartz substrate using a sub-100 µm stripe-patterned polydimethylsiloxane stamp for aligned carbon nanotube generation followed by fabrication of field effect transistor (FET). Atomic force microscopy, field emission scanning electron microscopy and Raman spectroscopy are employed to characterize the synthesized nanotubes. Unlike previous reports, the adopted approach enables direct electronic detection of bio-toxins with sensitivities comparable to ELISA. As a proof of concept, the fabricated FET responds to nM concentration levels (with a LOD of ∼2 nM) of epsilon toxin produced by Clostridium perfringens and a prominent food toxin. This facile approach could be customized to detect other classes of toxins and biomarkers upon appropriate functionalization of the aligned carbon nanotubes. Finally, we demonstrate the use of the FET-platform for detection of toxin in more complex matrices such as orange juice.


Subject(s)
Bacterial Toxins/analysis , Conductometry/instrumentation , Food Analysis/instrumentation , Food Contamination/analysis , Nanotubes, Carbon/chemistry , Nanotubes, Carbon/ultrastructure , Transistors, Electronic , Biosensing Techniques/instrumentation , Equipment Design , Equipment Failure Analysis , Staining and Labeling
2.
Acta Crystallogr D Biol Crystallogr ; 58(Pt 10 Pt 2): 1879-81, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12351845

ABSTRACT

Bucain is a three-finger toxin, structurally homologous to snake-venom muscarinic toxins, from the venom of the Malayan krait Bungarus candidus. These proteins have molecular masses of approximately 6000-8000 Da and encompass the potent curaremimetic neurotoxins which confer lethality to Elapidae and Hydrophidae venoms. Bucain was crystallized in two crystal forms by the hanging-drop vapour-diffusion technique in 0.1 M sodium citrate pH 5.6, 15% PEG 4000 and 0.15 M ammonium acetate. Form I crystals belong to the monoclinic system space group C2, with unit-cell parameters a = 93.73, b = 49.02, c = 74.09 A, beta = 111.32 degrees, and diffract to a nominal resolution of 1.61 A. Form II crystals also belong to the space group C2, with unit-cell parameters a = 165.04, b = 49.44, c = 127.60 A, beta = 125.55 degrees, and diffract to a nominal resolution of 2.78 A. The self-rotation function indicates the presence of four and eight molecules in the crystallographic asymmetric unit of the form I and form II crystals, respectively. Attempts to solve these structures by molecular-replacement methods have not been successful and a heavy-atom derivative search has been initiated.


Subject(s)
Bungarus , Elapid Venoms/chemistry , Elapid Venoms/toxicity , Toxins, Biological/chemistry , Toxins, Biological/toxicity , Amino Acid Sequence , Animals , Chromatography, Gel , Chromatography, High Pressure Liquid , Crystallization , Crystallography, X-Ray/methods , Elapid Venoms/isolation & purification , Molecular Sequence Data , Molecular Weight , Sequence Alignment , Sequence Homology, Amino Acid , Toxins, Biological/isolation & purification
3.
Acta Crystallogr D Biol Crystallogr ; 56 (Pt 12): 1702, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11092952

ABSTRACT

In the paper by Kuhn et al. [Acta Cryst. (2000), D56, 1401-1407] the name of the third author was given incorrectly. The correct name should be Doina-Silviana Comsa as given above.

4.
Acta Crystallogr D Biol Crystallogr ; 56(Pt 11): 1401-7, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11053837

ABSTRACT

Bucandin is a novel presynaptic neurotoxin isolated from Bungarus candidus (Malayan krait). It has the unique property of enhancing presynaptic acetylcholine release and represents a family of three-finger toxins with an additional disulfide in the first loop. There are no existing structures from this sub-category of three-finger toxins. The X-ray crystal structure of bucandin has been determined by the Shake-and-Bake direct-methods procedure. The resulting electron-density maps were of outstanding quality and allowed the automated tracing of 61 of the 63 amino-acid residues, including their side chains, and the placement of 48 solvent molecules. The 0.97 A resolution full-matrix least-squares refinement converged to a crystallographic R factor of 12.4% and the final model contains 118 solvent molecules. This is the highest resolution structure of any member of the three-finger toxin family and thus it can serve as the best model for other members of the family. Furthermore, the structure of this novel toxin will help in understanding its unique ability to enhance acetylcholine release. The unique structure resulting from the fifth disulfide bond residing in the first loop improves the understanding of other toxins with a similar arrangement of disulfide bonds.


Subject(s)
Elapid Venoms/chemistry , Neurotoxins/chemistry , Amino Acid Sequence , Animals , Bungarus , Crystallography, X-Ray , Molecular Sequence Data , Molecular Structure , Sequence Homology, Amino Acid
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