Development of analytical methods for human fluids by capillary electrophoresis
| Project/Area Number |
10672179
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Kumamoto University |
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Principal Investigator |
UJI Yoshinori Laboratory Medicine, Kumamoto University, Assistant Professor, 医学部, 助手 (90203512)
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| Co-Investigator(Kenkyū-buntansha) |
宇治 義則 熊本大学, 医学部, 助手 (90203512)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | capillary electrophoresis / LD isoenzymes / Alp isoenzymes / immunofixation by subtraction / serum protein analysis / アイソザイム分析 |
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| Research Abstract |
I have developed and evaluated the analytical method for human fluids by capillary eletrophoresis(CE). In the serum protein fractions, good linear correlation was observed between CE and cellulose acetate membrane electrophoresis (CA). The substantially higher values found for the α-1G fraction with a when compared with CA is related to the fact bothα-1 antitripsin and α-1 acidglycoprotein are quantitated by direct absorption with CE whereas with CA, the high sialic acid content of α-1-acid glycoprotein interferes with the binding of dyes used to quantitated the protein fraction. Reference ranges for albumin, α-1-G, and A/G were significantly different between CE and CA. No fiblinogen peak can be found in plasma sample with CE, because the concentration of fiblinogen is low in plasma samples. No interfering substances were observed both CA and CE. In the identification of serum M-protein by a immunofixation by subtraction (CE-IFEs), concordance studied between /IFE/s and agarose gel im
… More
munofixation eletrophoresis showed good agreement in identifying 30 monoclonal gammapathy patient samples. However, missed typing was observed containing very small monoclonal component or a second monoclonal hidden under a larger one or very close to one typed. Classical method, like immunoelectrophoresis or immunofixation, should be used for these samples. Separation and quantitative estimation of the isoenzymes of lactate dehydrogenase(LD) and alkaline phosphatase (ALP) isoenzyme in serum were accomplished. A uncoated fused sillica capillary column 50 cm(long) X 75mm( I.D. ) and substrate containing running buffer (L-lactic acid and NAD for LD isoenzymes,α-napthyl phospate and MgClィイD22ィエD2 for ALP isoenzymes) were used. The resulting product NADH for LD andα- napthol for ALP were detected at 340 nm. The results obtained by the proposed method both LD and ALP were correlated well with CA method. In conclusion, proposed analytical method by capillary eletrophoresis system is sensitive, precise and easy for clinical laboratory use. Less
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Report
(3 results)
Research Products
(10 results)
