Novel formaldehyde combined polypeptide and application thereof
A technology of formaldehyde and binding capacity, applied in the direction of carrier binding/immobilization of peptides, peptides, depsipeptides, etc., can solve problems such as damage to DNA damage repair mechanism, inability to completely remove, and failure.
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Embodiment 1
[0073] Example 1 The target magnetically loaded polypeptide has formaldehyde binding properties
[0074] If the target polypeptide (i.e. T1 or T2) has formaldehyde-binding properties, add a certain amount of target polypeptide to a specific formaldehyde system (such as formaldehyde aqueous solution) to make the two contact each other and fully combine for a period of time under suitable conditions Finally, the target polypeptide bound to formaldehyde is removed from the formaldehyde system. After sampling and measurement, the formaldehyde concentration in the formaldehyde system will inevitably decrease, so that the formaldehyde binding performance of the target polypeptide can be verified and explored. The specific process is as follows.
[0075]Entrust Shanghai Taopu Biotechnology Co., Ltd. to synthesize polypeptides T1 and T2 as shown in SEQ ID NO: 1 and 2, and the last amino acid phenylalanine at the C-terminal of these two polypeptides (the single-letter code for this amin...
Embodiment 2
[0084] Example 2 Saturation binding capacity of magnetically loaded polypeptides to formaldehyde
[0085] It can be known from Example 1 that the target polypeptide has a good formaldehyde binding capacity, however, the formaldehyde binding capacity per unit target polypeptide must be further explored to provide reference and basis for subsequent applications. Under specific systems and conditions, the formaldehyde binding capacity of the unit target polypeptide will increase with the prolongation of the reaction time until reaching and maintaining its saturated binding capacity. Therefore, this feature can be used to explore the saturated binding capacity of the target polypeptide to formaldehyde. The specific process is as follows .
[0086] Carry out the exploration of Example 2 with reference to Example 1 (a blank control group is also set up), the difference is that the reaction time of step (4) in Example 1 is only 30min, and the reaction time of Example 2 is extended to...
Embodiment 3
[0090] Example 3 The recovery of magnetic-carrying polypeptide and formaldehyde binding ability
[0091] After the target polypeptide achieves saturated binding to formaldehyde, in order to realize the reciprocal utilization of the target polypeptide for practical application, it is necessary to explore the conditions for its debinding after formaldehyde binding to restore its formaldehyde binding ability. The specific process is as follows.
[0092] For the magnetically loaded polypeptide system after 24 hours of reaction in Example 2 (at this time, the polypeptide has reached the saturated binding state of formaldehyde), use a magnetic frame to adsorb it on the side wall of the centrifuge tube, and use a pipette gun to remove as much as possible from the centrifuge tube. The reaction solution, and then wash the magnetic beads with pure water twice, then remove the pure water. Add 5ml of 0.15mol / L NaOH solution and 5ml of 0.3mol / L NaCl solution (at this time, the final concen...
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