Gd-DTPA-Polylysine-McAb junctional complex, preparation method and application thereof
A combination of monoclonal antibody technology, applied in the field of combination, can solve the problems of non-targeting, inability to accurately distinguish benign and malignant lesions, and difficulty in early diagnosis of diseases
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Embodiment 1
[0039] Preparation of a Gd-DTPA-Polylysine-McAb (anti-VEGF) conjugate
[0040] I. Synthesis of DTPA bis-anhydride (CaDTPA): 10g of diethylenetriaminepentacid (DTPA) was suspended in a mixture of 30ml of acetic anhydride and 15ml of pyridine, stirred and reacted at 60°C for 6h, after cooling, add an appropriate amount of ethanol to filter, and wash with ether , and dried to obtain a white solid DTPA bisanhydride (CaDTPA) crude product. After recrystallization, it is determined that the melting point of the product is 178~180°C (decomposition), and it has 1845, 1785 cm in the infrared spectrum. -1 C=O stretching vibration doublets of cyclic anhydrides.
[0041] II. Preparation of DTPA-Polysine: weigh Polylysine (polylysine) 10mg, dissolve in 3ml of 0.2mol / L NaH 2 CO 3 / Na 2 HCO 3 DTPA bis-anhydride (CaDTPA) solution was added in the buffer solution (PH=9.6) under constant stirring in an ice bath, and stirred at room temperature for 16 hours to complete the reaction. The re...
Embodiment 2
[0049] Experiment of Gd-DTPA-polylysine-McAb (anti-VEGF monoclonal antibody) in vivo MR molecular imaging of nude mouse model bearing human breast cancer
[0050] Materials and Methods
[0051] 1. Preparation of animal models
[0052] In this experiment, the cell suspension method was used to inoculate the human breast cancer cell line MDA-MB-231 in situ to make a nude mouse model of breast cancer. Nude mice and breast cancer MDA-MB-231 cell lines were purchased from the Cancer Institute of the Chinese Academy of Sciences. The 12 nude mice were all 5-week-old female specific pathogen-free (SPF) BALB / c nude mice, with a mass of 18-20 g. Breast cancer MDA-MB-231 cells were cultured with Leibovitz's L-15medimu (GIBCO) adding 15% fetal bovine serum (GIBCO). 7 / 0.2ml / only inoculated into the fat pad of the second pair of mammary glands on the right side of nude mice. After inoculation, the tumor growth was observed at least three times a week. After 2 weeks, a visible tumor was ...
Embodiment 3
[0075] Control experiment of Gd-DTPA-polylysine-McAb (anti-VEGF monoclonal antibody) and Gd-DTPA in MR molecular imaging of nude mouse model of breast cancer
[0076] At present, the incidence of breast cancer at home and abroad is showing a significant upward trend. The incidence of female breast cancer in many large cities in my country has ranked first in female malignant tumors, and the age of onset tends to be earlier. The diagnosis of breast cancer mainly relies on imaging examinations, including X-ray, B-ultrasound and MRI, etc. Among them, high-field-strength MRI dynamic multi-phase enhanced scanning has the greatest value, and its specificity and sensitivity are high. When extensive Gd-DTPA is present, the MRI findings of breast benign and malignant tumors overlap and cross each other. Therefore, specific diagnosis cannot be achieved, which not only affects the identification of breast cancer and benign lesions, but also affects the detection of early breast cancer. ...
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