Differential antigenic protein recovery from Taenia solium cyst tissues using several detergents.
Human and porcine cysticercosis is caused by the larval stage of the flatworm Taenia solium (Cestoda). The protein extracts of T. solium cysts are complex mixtures including cyst's and host proteins. Little is known about the influence of using different detergents in the efficiency of solubilization-extraction of these proteins, including relevant antigens. Here, we describe the use of CHAPS, ASB-14 and Triton X-100, alone or in combination in the extraction buffers, as a strategy to notably increase the recovery of proteins that are usually left aside in insoluble fractions of cysts. Using buffer with CHAPS alone, 315 protein spots were detected through 2D-PAGE. A total of 255 and 258 spots were detected using buffers with Triton X-100 or ASB-14, respectively. More protein spots were detected when detergents were combined, i.e., 2% CHAPS, 1% Triton X-100 and 1% ASB-14 allowed detection of up to 368 spots. Our results indicated that insoluble fractions of T. solium cysts were rich in antigens, including several glycoproteins that were sensitive to metaperiodate treatment. Host proteins, a common component in protein extracts of cysts, were present in larger amounts in soluble than insoluble fractions of cysts proteins. Finally, antigens present in the insoluble fraction were more appropriate as a source of antigens for diagnostic procedures.
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico.
José
J
Navarrete-Perea
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico.
Rodrigo
R
Orozco-Ramírez
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico.
Bárbara
B
Moguel
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico.
Edda
E
Sciutto
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico.
Raúl J
RJ
Bobes
Department of Immunology, Institute for Biomedical Research, Universidad Nacional Autónoma de México, Mexico D.F. 04510, Mexico. Electronic address: laclette@biomedicas.unam.mx.
Juan P
JP
Laclette
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D001622:Betaine / Q000031:analogs & derivatives / Q000737:chemistry; D002021:Buffers; D002793:Cholic Acids / Q000737:chemistry; D003560:Cysts / Q000737:chemistry* / Q000276:immunology / Q000469:parasitology; D003902:Detergents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006023:Glycoproteins / Q000302:isolation & purification; D006801:Humans; D018482:Muscle, Skeletal / Q000469:parasitology; D013552:Swine; D041201:Taenia solium / Q000737:chemistry* / Q000276:immunology; D013622:Taeniasis / Q000469:parasitology
D000947:Antigens, Helminth; D002021:Buffers; D002793:Cholic Acids; D003902:Detergents; D006023:Glycoproteins; C506587:amidosulfobetaine-14; D001622:Betaine; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate
ASB-14; CHAPS; Cysticercosis; Triton X-100