Device and method for capturing specificity of circulating cancer cells in peripheral blood
A technology for circulating cancer cells and peripheral blood, which is applied to tumor/cancer cells, biological material sampling methods, animal cells, etc. It can solve the problems of long pretreatment of sample cells, failure to meet CTC, and lack of specific antibodies, etc., to reduce Effects of light scattering, improved signal-to-background ratio, and large liquid flux
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Embodiment 1
[0035] Such as figure 1 with figure 2 As shown, the device for cell-specific capture includes a syringe 1, a square capillary 3 communicated with the syringe 1 through a Teflon capillary 2; the inner wall of the square capillary 3 is adsorbed with avidin 6 and biotin-aptamer Complex 7.
Embodiment 2
[0036] Example 2: Specific capture method of circulating liver cancer cells in peripheral blood
[0037] (1) Synthesis of DNA sequence L1
[0038] Nucleic acid aptamer probe L1 (the italic part is the nucleic acid sequence that recognizes liver cancer cells, and the 3' end is modified with biotin):
[0039] 5'- AGTCCATTTTATTCCTGAATATTTGTTAACCTCATGGAC TTTTTTTTTT-3'-biotin
[0040]The DNA sequence L1 used to capture liver cancer cells was synthesized using an ABI3400 DNA / RNA synthesizer. Synthetic starting materials were purchased from Glen Research (Sterling, VA). DNA purification was achieved by ProStar HPLC (Varian). DNA concentration was determined by Cary Bio-300 UV-visible spectrometer (Varian).
[0041] (2) Immobilization of DNA sequence L1 on the capture device
[0042] Wash and activate the square capillary with 50 uL 1M NaOH; wash 5 times with 50 uL deionized water, then introduce 50 uL D-PBS containing 5 mg / mL avidin into the treated square capillary with a s...
Embodiment 3
[0048] Example 3: Specific capture method for circulating small cell lung cancer cells (SCLC) in peripheral blood
[0049] (1) Synthesis of DNA sequence L2
[0050] Nucleic acid aptamer probe L2 (the italic part is the nucleic acid sequence that recognizes small lung cancer cells, and the 3' end is modified with biotin):
[0051] 5'- GTGGATTGTTGTGTTCTGTTGGTTTTTGTGTTGTG TTTTTTTTTT-3'-biotin;
[0052] The DNA sequence L2 used to capture small lung cancer cells was synthesized using an ABI3400 DNA / RNA synthesizer. Synthetic starting materials were purchased from Glen Research (Sterling, VA). DNA purification was achieved by ProStar HPLC (Varian). DNA concentration was determined by Cary Bio-300 UV-visible spectrometer (Varian).
[0053] (2) Immobilization of DNA sequence L2 on the capture device
[0054] Wash and activate the square capillary with 50 uL 1M NaOH; wash 5 times with 50 uL deionized water, then introduce 50 uL D-PBS containing 5 mg / mL avidin into the treated...
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