Tumor-targeting fusion protein drug carriers for optical imaging and photodynamic therapy
A fusion protein and drug technology, applied in the field of biomedicine, can solve the problem of poor targeting selectivity of water-soluble tumors
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[0076] Construction, expression, purification and characterization of ATF-HSA fusion protein drug carrier:
[0077] The ATF-HSA fusion protein gene (SEQ ID NO.2) pPICZαA expression vector (purchased from Invitrogen, USA) was constructed by gene cloning. After identification by gene sequencing, the expression vector was electrotransformed into competent cells of Pichia pastoris (Pichia pastoris) X-33 (purchased from Invitrogen, USA). After a small amount of expression was identified, the fusion protein drug carrier was expressed in large quantities in the eukaryotic expression system, and methanol was added every 24 hours to a final concentration of 1%. After induction of expression for 4 days, centrifuge at 4°C, 7000rpm, 10min to remove the bacteria, and use sodium acetate buffer (AB) to adjust the pH to pH4.5. Centrifuge at 4°C, 10000rpm for 40min, and vacuum filter through a 0.45μm, 0.22μm filter membrane Finally, the treated mass-induced expression culture solution was dil...
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