1989 Fiscal Year Final Research Report Summary
Neuropathological Diagnosis of Pulp Disease Using Antimyelin-protein Monoclonal Antibody
| Project/Area Number |
63480422
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Conservative dentistry
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| Research Institution | Kyushu University |
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Principal Investigator |
NAGASAWA Hisashi Kyushu University, Faculty of Dentistry, Professor, 歯学部, 教授 (10013848)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Yoshihiko Kyushu University, Faculty of Dentistry, Assistant Professor, 歯学部, 講師 (20150477)
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| Project Period (FY) |
1988 – 1989
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| Keywords | Peripheral Nerve / Myelin Glycoprotein / Monoclonal Antibody / Neuropathological Diagnosis / Pulp |
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| Research Abstract |
The P0 protein is an integral membrane glycoprotein which comprises 50-60% of the total myelin proteins in mammalian peripheral nerves. Then, in the neuropathological examination of pulp diseases, this glycoprotein could be used as the marker. This study was designed to isolate and purify P0 glycoprotein from a small material. The myelin fractions were prepared from axillar, musculocutaneous, median, radial, ulnar and sciatic nerves of rats by sucrose density gradient centrifugation. This myelin fractions were examined with electron microscope. The myelin fraction consisted mainly of the lammellar structure. Subcellular organelles, such as nuclei and mito chondria, were not observed in this fraction. The myelin fraction was homogenized in 0.03 M HC1, stirred at 4ﾟC and centrifuged. The acid-insoluble residue was homogenized directly with chloroform/reethanol. The fat-free precipitate was dissolved in 0.1 M phosphate buffer containing 10% sodium dodecyl sulfate (SDS). The supernatant was applied on a Sephacryl S-100 column, equilibrated and eluted with 0.1 M phosphate buffer containing 2% SDS. One major peak was detected. The fractions from major peak was concentrated. This major peak showed a single band (about 32 KD) when the gel was stained by Coomassie blue for proteins and periodic acid-Schiff reagent for glycoproteins after SDS-polyacrylamide gel electrophoresis. These findings suggest that the isolated and purified protein in the present study is P0 glycoprotein and the concentrated sample could be directly used as the antigen for preparing monoclonal antibody against PO glycoprotein.
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