A kind of synthesis method and application of polydopamine-modified maldi target plate
A polydopamine and synthesis method technology, applied in advanced nanomaterials and nanometer fields, can solve the problems of mass spectrometry ion source pollution, damage, etc., to avoid cumbersome elution, simple synthesis method, good biocompatibility and hydrophilicity sexual effect
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Embodiment 1
[0029] Synthesis of polydopamine-modified MALDI target plate
[0030] (1) Disperse 400mg of dopamine hydrochloride in 200mL of Tris buffer (10mM, pH=8.5).
[0031] (2) Wash the MALDI target plate several times with distilled water and ethanol, and dry at room temperature.
[0032] (3) Submerge the washed MALDI target plate in the dopamine solution in (1) and react at room temperature for 24 hours.
[0033] (4) The polydopamine-modified target plate obtained in (3) was washed three times with distilled water, and immersed in Ti(SO 4 ) 2 (100mM) aqueous solution at room temperature for 2 hours.
[0034] (5) Rinse the target plate obtained in (4) three times with distilled water and dry at room temperature.
[0035] figure 1 It is a scanning electron micrograph of the surface of the MALDI target plate modified by polydopamine. Dopamine presents a small spherical shape at the beginning of the polymerization reaction, and tends to form a polydopamine film at the later stage o...
Embodiment 2
[0036] Example 2: Polydopamine-modified MALDI target plate selectively enriches phosphorylated peptides and performs high-throughput MALDI-TOFMS analysis
[0037] (1) Sample preparation: Dissolve BSA or β-casein protein in NH 4 HCO 3 Buffer (25mM, pH8.3) and digested with trypsin (2%, w / w) at 37°C for 16 hours. The product after enzymatic hydrolysis should be stored below 0°C. After the serum samples were centrifuged, the supernatant was stored below 0°C.
[0038] (2) Spotting the target: wash the modified target plate with 50% acetonitrile and 0.1% TFA aqueous solution (v / v) three times, respectively add the β-casein enzymatic hydrolysis solution, the mixture of β-casein and BSA enzymatic hydrolysis solution and The treated serum was diluted to different concentrations with 50% acetonitrile and 0.1% TFA aqueous solution (v / v); pipette 1 μL of the diluted enzymolysis solution onto the modified MALDI target plate, and place it in a wet box at room temperature for enrichment ...
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