JP2005245381A - 免疫細胞クローンの拡大の有無の判定方法 - Google Patents
免疫細胞クローンの拡大の有無の判定方法 Download PDFInfo
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Abstract
【解決手段】 抗原に応答した免疫細胞クローンの拡大の有無の判定方法は、生体から採取された免疫細胞の抗原レセプター遺伝子群又は免疫グロブリン遺伝子群中の、多様性の高い領域の隣接領域にプライマーを設定し、多様性の高い領域を鋳型として、ダイデオキシ法による伸長反応を行なう工程と、前記伸長反応により生成された核酸断片を質量分析にかける工程と、質量分析の結果から、特定の免疫細胞クローンの拡大が起きたか否かを判定する工程とを含む。
【選択図】 図1
Description
Vhの配列の一部に相補的な配列
cagcttcaggagtcagg
cagctgaagcagtcagg
Vkの配列の一部に相補的な配列
attgtgatgacacagtctcc
gttgatgacccaaact
その他
tgtgatattgtgctaactcagtct
gggccctctgggctcaatttt
Jbの配列の一部に相補的な配列
actgtgagtctggttggtttacc
cctggtccctgacgggaag
Vbの配列の一部に相補的な配列
ctgaatgcccagacagctccaagc
cattattcatatggtgctggc
その他
tctgcagcctgggaatcacaa
ctgctaagaaaccatgtacca
合成鋳型DNAである#59(59 mer)の塩基配列
gcc agc agc ccc ggg aca gac aca gaa gtc ttc ttt ggt aaa gga acc aga ctc aca gt
マトリックス用3-ヒドロキシピコリニックアシッド(3-hydroxypicolinic acid for a matrix)
ポジティブモード、リニアモード
イオン源1: 20.1kV, イオン源2: 18.7 kV
鋳型として用いるH1.110(62mer)の塩基配列
gcc agc ggt gag aga cac cta aac aca gaa gtc ttc ttt ggt aaa gga acc aga ctc aca gt
act gtg agt ctg gtt cct tta cc
Claims (5)
- 生体から採取された免疫細胞の抗原レセプター遺伝子群又は免疫グロブリン遺伝子群中の、多様性の高い領域の隣接領域にプライマーを設定し、多様性の高い領域を鋳型として、ダイデオキシ法による核酸の伸長反応を行なう工程と、前記伸長反応により生成された核酸断片を質量分析にかける工程と、質量分析の結果から、特定の免疫細胞クローンの拡大が起きたか否かを判定する工程とを含む、抗原に応答した免疫細胞クローンの拡大の有無の判定方法。
- 前記免疫細胞がT細胞又はB細胞である、請求項1記載の方法。
- 前記多様性の高い領域がCDR領域である請求項1又は2記載の方法。
- 前記免疫細胞がT細胞であり、前記多様性の高い領域がレセプターβ鎖CDR3領域である請求項3記載の方法。
- 前記プライマーを設定する領域が、V領域の下流部分又はJ領域の上流部分である請求項1ないし4のいずれか1項に記載の方法。
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