研究業績集
学会発表2013年
タイトル
Novel sandwich immunoassay for quantification of 25-hydroxy vitamin D in human blood
発表者
K. Omi, T. Ando, T. Sakyu, H. Sato, A. Oka, Y. Hotta, M. Horiike, T. Shirakawa, K. Goishi
学会名
AACC's 45th Annual Oak Ridge Conference
内容
Vitamin D is well known for its role as an important regulator of calcium homeostasis and bone remodeling, and also known to affect the development of several non-bone diseases. In spite of the clinical
importance, the available automated competitive immunoassays reportedly fail to meet accuracy and specificity requirements.
Here we show a novel sandwich immunoassay for 25OH-D using an antibody
that specifically recognizes an immunocomplex consisting of 25OH-D and anti-25OH-D antibody. The assay, developed on a fully automated chemiluminescence analyzer LUMIPULSE G1200, specifically detected
25OH-D immunocomplex in a dose-dependent manner, and it well correlated with the commercially available RIA assay (r >0.98). Total assay CVs for 14 ng/mL, 39 ng/mL, and 97 ng/mL samples were 2.2%, 1.9%,
and 3.3%, respectively. Using two antibodies, our assay showed improved specificity against immunoreactive derivatives such as 24,25(OH)2-D, which can be present in human serum and known to cross-react
with antibodies used in most commercially available immunoassay kits.
Our novel sandwich immunoassay platform would provide high-throughput, accurate and specific immunoassay for 25OH-D, leading
to the breakthrough in routinely performed 25OH-D assay.
タイトル
Novel chemiluminesence immunoassay platform for small molecules using immuno-complex antibodies: proof of concept in E2, FT3 and Vitamin D assay with high sensitivity and specificity
発表者
T. Sakyu, T. Ando, A. Oka, F. Takemura, T. Shirakawa, K. Goishi, K. Omi
学会名
AACC's 45th Annual Oak Ridge Conference
内容
Small molecules immunochemically classified as haptens are measured with competitive immunoassay, which is theoretically inferior to noncompetitive sandwich immunoassay. Although some reports have been made
on the sandwich immunoassay for haptens using antibodies that react with hapten-antibody complex, it is virtually impossible to obtain such antibodies systemically by conventional methods. Moreover,
no automated assays for haptens using such antibodies have been reported so far.
Here we demonstrate a novel sandwich immunoassay format for haptens using antibodies specifically recognizing hapten-antibody
immunocomplex, which we established with ADLib system, an ex vivo antibody developing system. Using the established antibodies, we have successfully established sandwich immunoassays for haptens including
E2, T3 and 25-OH D. Furthermore, these assays were applicable to LUMIPULSE G1200.
These results suggest that our platform enables the systematic establishment of high-throughput sandwich immunoassays
for haptens. Our hapten sandwich immunoassay platform should be the simplest and most practical approach for routine assays of haptens leading to the breakthrough in analytical/clinical chemistry.
タイトル
Novel sandwich immunoassay for quantification of 25-hydroxyvitamin D on fully automated analyzer
発表者
T. Ando, Y. Kitamura, T. Sakyu, Y. Uchida, M. Tomita, F. Takemura, T. Shirakawa, K. Aoyagi, K. Goishi and K. Omi
学会名
AACC Annual Meeting & Clinical Lab Expo 2013
内容
Vitamin D plays important roles in regulating calcium homeostasis and bone remodeling, and association between vitamin D insufficiency and several non-bone diseases have also been reported. 25-hydroxyvitamin
D (25OH-D) is considered to be the best vitamin D marker, and 25OH-D can be measured by competitive immunoassay, HPLC and LC-MS/MS. As the number of tests increases, demand for an automated 25OH-D assay
continues to grow.
In this study, we successfully established sandwich immunoassay using an anti-immunocomplex antibody specifically recognizing an immunocomplex consisting of 25OH-D and anti-25OH-D
antibody. The sandwich assay developed on a fully automated chemiluminescence analyzer, LUMIPULSE G1200 showed improved cross-reactivity with derivatives and significant correlation with the commercially
available Total Vitamin D RIA assay (r >0.98).
Our novel sandwich immunoassay platform would provide high-throughput, accurate and specific immunoassay for haptens including hormones, vitamins, drugs
and toxins, leading to the breakthrough in analytical/clinical chemistry.
タイトル
Cisplatin暴露は上皮間葉移行様変化を誘導し分子標的薬に対する耐性獲得に影響を及ぼす
発表者
黒川 真季1), 伊勢 伸之, 小見 和也, 碁石 勝利, 東山 繁樹1,2)
1) Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine
2) Division of Cell Growth
and Tumor Regulation, Proteo-Science Center
学会名
第86回日本生化学会大会
内容
プラチナ製剤等細胞障害性抗癌剤及び分子標的薬の開発により抗癌剤治療が劇的に進化しつつあるが、治療過程で常に薬剤耐性が誘導されることが治療上の限界となる。臨床においてはこれら異なる二つのタイプの薬剤を連続して使用するケースがあるものの、一方の薬剤により誘導された耐性が他方の薬剤に対する感受性・耐性に与える影響に関しては殆ど解明されていない。本研究では、肺癌細胞株を用いたcisplatin、分子標的薬連続治療モデル系を構築し、プラチナ製剤治療が分子標的薬治療に与える影響を検討した。また、双方の薬剤に共通する耐性機構である上皮間葉移行が、本連続治療モデルに関与する可能性を併せて検討した。
