| Project/Area Number |
18590534
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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 | Sapporo Medical University |
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Principal Investigator |
TSUJI Naoki Sapporo Medical University, SCHOOL OF MEDICINE, ASSISTANT PROFESSOR (00347171)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Naoki SAPPORO MEDICAL UNIVERSITY, SCHOOL OF MEDICINE, Professor (10158644)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,860,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | ANTI-ANIMAL ANTIBODY / HAMA / ELISA FOR DETECTIONOF HAMA / HETEROPHILIC BLOCKING REAGENT |
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| Research Abstract |
In the present study, we developed enzyme-linked immunosorvent assay (ELISA) for the detection of human anti-mouse antibodies (HAMA) in the serum and investigate the frequency of people possesses HAMA in Japan.
34 cases out of 290 cases (11.7%) were positive (IgG type ; 18 cases, IgM type 20 cases). The most frequent IgG subtype that HAMA recognized was IgG1 (IgG type ; 14 cases, IgM type 11 cases). In addition, some cases showed positive against two or three kinds of IgG subtype (IgG type ; IgG2a+IgG2b:3 rases, IgG1+IgG2a+IgG2b:1ca.p, IgM type ; IgG2a+IgG2b:5 cases, IgG1+IgG2a + IgG2b:2 case).
To eliminate the effect of HAMA on the immunoassays, heterophilic blocking reagent (HBR) which contains mouse immunoglobulins is added to sample or assay reagents. We then determined the effect of HBR on HAMA using ETISA system. When 10 or 15μl of HBR was added to 100 ul of sample, measured values were decreased to 30-50% of untreated samples. In contrast, when of HBR was added to sample, measured values were significantly increased 2-3 times of untreated samples. These results indicate that establishing the amount of HBR added to samples is important to block the effect of HAMA on the immunoassays.
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