高感度検出技術であるイムノPCR

Technology, Information, and Service for Life Science　　CareTIS, Co., Ltd.

Immuno-PCR, a high-sensitivity antigen detection technology

【Technical content】
	Immuno-PCR was first reported by Sano et al. In 1992 as a new antigen detection method that combines the specificity of the antigen-antibody reaction with the high sensitivity of the PCR method. For more information, please refer to http://www.jpo.go.jp/shiryou/s_sonota/hyoujun_gijyutsu/kakusan/0056.html
The problem is；
	In the case of the antigen immobilization method, it is diffcult to use becouse the detection concentration is low for detecting antigens that are present only in trace amounts such as blood markers and cultured cells. In the sandwich method, there is a possibility that the biotinylated secondary antibody and streptavidin bind nonspecifically. As a result, the DNA will be amplified even if it dose not contain an antigen, and it will be detected as a peak. At CareTIS, we can offer by improoving the blocking agent, and the nucleic acid (tag) that binds to the secondary antibody.

Next, We would like to introduce an example of immuno-PCR performed at CareTIS.

A：Examination of the sensitivity of immuno-PCR method (sandwich method).

【Summary】
Fluorescein-labeled BSA (homemade product:F-BSA) was deected using the immuno-PCR method (sandwich method). Higher sensitivity detection is expected by improving solid phase blocking agents.

【Keywords for materials and method】
・F-BSA (antigen)　　　　　　　　　・Top Yield (solid phase plate:Nunc)
・Anti-BSA Ab (primary antibody) 　　　　・Agilent2100 (measuring device)
・Fluorescein Ab(secondary antibody)　・PCR（20 cycles）

【Example】
	Measurement system
	Agilent 2100　　　　　　　　　　　　　　　　　　Thermal Cycler
	Molecure binding in the sandwich method

	Detection chart

Shows the PCR product, and it can be seen that it is proportional to the amount of antigen (F-BSA).
	Detection result
	If you woule like to know more details, please contact us by email.

Ｂ： Examination of the sensititivity of immuno-PCR (antigen direct immobilization method)

【Sammary】
Florescein-labeled BSA (homemade product : F-BSA) was detected using the immuno-PCR method (antigen immobilization). Here, only 0.2ng/μl was applied, but the sensitivity was almost the same as that of the sandwich method. Higher sensitivity detection is expected by involving solid phase blocking agents.

【Keywords for materials and method】
・F-BSA (antigen)　　　　　　　　　・Top Yield (solid phase plate:Nunc)
・Anti-BSA Ab(antibody)）　　　　　　　・Agilent2100 (measuring device)
・PCR (20 cycles)

【Example】
	Molecule binding in antigen immobilization method

	Detection chart

	Shows the PCR product, and it can be seen that the sensitivity is almost the same as that of the sandwich method, and that it is proportional to the amount of antigen (F-BSA).