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Preparation of an infectious mononucleosis receptor from sheep erythrocyte stroma
Abstract:
A method is described for the isolation and purification of infectious mononucleosis receptor (IMR) in high yield from sheep erythrocyte stroma. The procedure involves extraction of the stroma successively with boiling acetone, 100, 75 and 50% ethanol which removes approximately 40% non-infectious mononucleosis-active material. Subsequent phenol/buffered saline extraction of the residue from the organic extraction yields a highly active IMR preparation in the aqueous phases. This material was further purified by ethanol precipitation and gel filtration chromatography. The yield was 0.2% of the stroma. Approximately 0.1 mug/ml of the purified IMR inhibits 4 agglutinating units of infectious mononucleosis (IM) serum. The material is a glycoprotein containing N-acetyl- and N-glycolylneuraminic acids, hexose, hexosamine and approximately 46% protein. It gives one band in immunoelectrophoresis with IM serum and loses IM specificity upon treatment with neuraminidase.
Insights
Researchers developed a high-yield method to isolate the infectious mononucleosis receptor (IMR) from sheep red blood cells. This purified glycoprotein is crucial for understanding infectious mononucleosis (IM) serum interactions.
Area of Science:
- Biochemistry
- Immunology
- Cell Biology
Background:
- Infectious mononucleosis (IM) is a common viral illness.
- The infectious mononucleosis receptor (IMR) plays a key role in the disease's pathogenesis.
- Efficient isolation of IMR is essential for further research.
Purpose of the Study:
- To develop a high-yield method for isolating and purifying the infectious mononucleosis receptor (IMR).
- To characterize the biochemical properties of the purified IMR.
- To assess the biological activity of the purified IMR.
Main Methods:
- Sheep erythrocyte stroma extraction using organic solvents (acetone, ethanol).
- Phenol/buffered saline extraction to isolate IMR.
- Purification via ethanol precipitation and gel filtration chromatography.
- Biochemical analysis including protein and carbohydrate content determination.
- Immunoelectrophoresis and neuraminidase treatment to assess specificity.
Main Results:
- A novel, high-yield purification method for IMR was established.
- The purified IMR is a glycoprotein containing specific amino and carboxyl sugars.
- The purified IMR demonstrated biological activity, inhibiting IM serum agglutination.
- IMR specificity was confirmed by immunoelectrophoresis and lost upon neuraminidase treatment.
Conclusions:
- The developed method provides a high yield of purified IMR.
- The purified IMR is a glycoprotein with specific biochemical and immunological properties.
- This purified IMR is a valuable tool for studying infectious mononucleosis.