Tumor-targeting coxsackie virus/adenovirus mimic peptide and application thereof
A coxsackie virus and peptide-mimicking technology, which is applied in the field of biomedical engineering, can solve many problems, and achieve the effect of convenient access and low synthesis cost
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Embodiment 1
[0061] 1. Labeling fluorescent dyes for in vitro experiments
[0062] The fluorescent dye used in the in vitro cell experiment of the present invention is Rhodamine B (Rhodamine B), referred to as RhB, respectively connected with the YQC-X (X=3-9) peptide of the present invention through an amide bond, referred to as YQC-X-RhB respectively , the connection method is specifically to take 1 mg rhodamine B (see references for details: Cao, J., et al., Fast clearing RGD-based near-infrared fluorescent probes for in vivo tumor diagnosis. Contrast Media Mol Imaging, 2012.7 (4): p.390-402) was dissolved in 1ml PBS (pH 7.4), added 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, N-hydroxysuccinimide ( EDC / NHS) (molar ratio Rhodamine B:EDC:NHS=1:1.5:1.5), react in the dark for 4 hours, and activate. 1 mmol YQC-X peptide was weighed and dissolved in 1 ml PBS buffer (pH 7.4) containing fluorescent dye Rhodamine B, and stirred overnight at room temperature in the dark. After ...
Embodiment 2
[0066] 1. Labeling fluorescent dyes for in vitro experiments
[0067] The fluorescent dye used in the in vivo animal experiment of the present invention is a near-infrared organic dye ICG-Der-02, which is respectively connected with YQC-X (X=3-9) through an amide bond, referred to as ICG-Der-02-C1, ICG-Der -02-C2, the specific method of connection is to take 2mg of ICG-Der-02 (see references for details) dissolved in 1ml of PBS (pH 7.4), add 1-ethyl-(3-dimethylaminopropyl) carbon Imide hydrochloride, N-hydroxysuccinimide (EDC / NHS) (molar ratio ICG-Der-02:EDC:NHS=1:1.5:1.5), protected from light for 4 hours, activated. Weigh 1 mmol of YQC-X (X=1-9) peptide and dissolve it in 1 ml of PBS buffer (pH=7.4) containing fluorescent dye ICG-Der-02, and stir overnight at room temperature in the dark. After the reaction, the reaction solution was concentrated and passed through a Sephadex G-25 column with PBS buffer (pH=7.4) as the eluent to obtain purified ICG-Der-02-C1, ICG-Der-02-C2 ...
Embodiment 3
[0073] The YQC-X (X=3-9) polypeptide found in the present invention is designed and developed through computer simulation, synthesized and purified, and the purity of the polypeptide is up to 98% as detected by high performance liquid chromatography. The Iodogen oxidation method can iodine-label the polypeptides containing tyrosine, histidine, and tryptophan residues, and the YQC-X (X=3-9) peptide sequence has histidine residues, so it can be Use this method for iodine labeling. The following uses YQC-5 as an example to describe the application.
[0074] 1. Polypeptide solid-phase synthesis
[0075] A: Coupling
[0076] First, swell 0.33 / 2mmol 6.06g of Fmoc-Lys-Rink amide MBHA resin (Fmoc-Lysine-rink amino resin) in dimethylformamide, 10mL / g resin, and then use 20% hexahydro Pyridine / dimethylformamide solution removes the Fmoc protecting group (hereinafter referred to as decapping), washes the resin after decapping with dimethylformamide, adds raw material Fmoc-Ala(Boc)-OH ...
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