ABSTRACT
<p><b>OBJECTIVE</b>To prepare and characterize rabbit polyclonal antibodies against Toxoplasma gondii vacuolar proton pyrophosphatase type I (TgVP1).</p><p><b>METHODS AND RESULTS</b>Two synthesized peptides TgVP1-1 and TgVP1-2 as the haptens were conjugated with KLH to immunize rabbits. Indirect ELISA showed that the titers of rabbit anti-TgVP1-1 polyclonal antibody and rabbit anti-TgVP1-2 polyclonal antibody reached 1:128 000. Western blotting results revealed that both purified polyclonal antibodies could specifically bind to a purified 85 kD T. gondii protein predicted as TgVP1. The protein detected by these two polyclonal antibodies was distributed in the cytoplasm of T. gondii tachyzoite, and this distribution pattern was consistent with that of acidocalcisome.</p><p><b>CONCLUSION</b>The peptide-based method of antibody generation is efficient and the obtained TgVP1 polyclonal antibodies possess a high specificity to facilitate further study of T. gondii acidocalcisome and the diagnosis of toxoplasmosis.</p>
Subject(s)
Animals , Rabbits , Antibodies , Allergy and Immunology , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Protozoan Proteins , Allergy and Immunology , Pyrophosphatases , Allergy and Immunology , ToxoplasmaABSTRACT
Using a novel hybrid technology combined differential ion mobility spectrometry ( DMS) with drift time ion mobility spectrometry DMS-IMS2 , we detected the typical chemical warfare agent simulants dimethyl methylphosphonate ( DMMP ) and methyl salicylate ( MS) . With carrier gas 800 mL/min and DMS RF voltage 1100 V, the chemical warfare agents DMMP and MS could be detected and characterized by DMS-IMS2 under DIMS mode. In addition, DMS-IMS2 is capable to monitor positive and negative ions of DMMP and MS simultaneously, and provides the two-dimensional separation parameters DMS compensation voltage ( CV) and IMS drift time ( Td ) , which provides more information for the identification of two chemical warfare agents. DMS-IMS2 has potential application in on-site detection and instrument miniaturization due to its advantages including small size, low power consumption and rapid response time.
ABSTRACT
In this article, an acetone-enhanced negative photoionization (AENP) source based on a 10. 6 eV vacuum ultraviolet ( VUV) lamp was developed and coupled to a home-made time-of-flight mass spectrometer for rapid detection of trace explosives. In the AENP source, acetone molecules absorbed 10. 6 eV photons and were ionized by single photon ionization to emit photoelectrons. The photoelectrons reacted with O2 , CO2 , etc. in the atmosphere to produce mainly CO-3 negative reactant ions. With this ionization source, common explosives, N-nitrobiz ( 2-hyolorolroxy ethyl )-amine dinitrate ( DINA ) , Tetryl, trinitrotoluene ( TNT ) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), could be detected sensitively, and the limit of detection of 2 pg ( TNT) with a linear range of 3 orders of magnitude was achieved. The simple structure, high sensitivity characteristics make the AENP source as a promising ionization source for mass spectrometry.