Trichinella spiralis: Proteins and antigens isolated from a large-particle fraction derived from the muscle larva |
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Authors: | Dickson D Despommier Anthony Laccetti |
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Institution: | Division of Tropical Medicine, School of Public Health, College of Physicians and Surgeons, Columbia University, 630 West 168 Street, New York, New York 10032, U.S.A. |
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Abstract: | Proteins and antigens derived from a large-particle fraction of muscle larvae of Trichinella spiralis (i.e., the S3 fraction) were characterized in terms of their molecular weights, isoelectric points, carbohydrate contents, electrophoretic mobilities, antigenicity, and their ability to induce protection in mice. Gel filtration on Sephacryl S-200 yielded 5 major peaks of material while electrophoresis in polyacrylamide gel with sodium dodecyl sulfate revealed a minimum of 28 proteins ranging in MW from 11,000 to 200,000. Analytical isoelectric focusing on acrylamide gel yielded 37 bands of protein, while the periodic acid-Schiff reaction performed on a similar gel revealed 22 glycoproteins. Most proteins were within a pI range of 4.0–7.0, while all of the glycoproteins had pI ranging from 4.0 to 6.5. Immunoelectrophoresis of the S3 fraction using hyperimmune rabbit serum demonstrated a minimum of 19 precipitin arcs, while crossed immunoelectrophoresis yielded 16 peaks. These determinations were made on several batches of material isolated in the same fashion and gave the same results. Preparative isoelectric focusing yielded 30 fractions. These fractions were assayed for the presence of antigens, then pooled and tested for their ability to induce protection in mice against an oral challenge infection. Fused rocket immunoelectrophoresis of all 30 fractions revealed the presence of a minimum of 18 antigens with pI ranging from 4.0 to 9.0. The pooled fractions (i.e., 1–9; 10–20; 21–30) all protected mice against oral challenge infection, while fraction 5 (pI = 4.3) protected best. |
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Keywords: | Nematode parasitic Cell fractionation Large-particle fraction Proteins Glycoproteins Electrophoresis Immunoelectrophoresis and crossed immunoelectrophoresis Molecular sizing Column chromatography Analytical isoelectric focusing Preparative flatbed isoelectric focusing Fused rocket immunoelectrophoresis Protection tests Protection-inducing antigens Mouse |
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