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目的研究抗P-糖蛋白(P-gp)的鼠源性单克隆抗体PHMA02在移植有K562/A02细胞瘤的裸鼠体内的影像分布。方法通过杂交瘤技术制备PHMA02抗体,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和Western-blot技术鉴定纯度,荧光激活细胞分选术(FACS)测抗体与K562和K562/A02细胞的结合活性,改良氯胺T法125I标记抗体,尾静脉注射入裸鼠,分时间点行SPECT扫描。结果纯化所得PHMA02纯度较高,与P-gp+的K562/A02细胞特异性结合阳性率为94.47%,注入125I标记的PHMA02后,P-gp+(K562/A02)组第2日全部裸鼠瘤区出现放射性浓聚,并在随后数日浓集进一步增强。结论125I-PHMA02抗体在P-gp+(K562/A02)组和P-gp-(K562)对照组的影像学差异可以明确区分P-gp+的肿瘤组织。
Objective To study the image distribution of PHMA02, a murine monoclonal antibody against P-glycoprotein (P-gp), in nude mice transplanted with K562 / A02 cell tumor. Methods PHMA02 antibody was prepared by hybridoma technique. Purity was determined by sodium dodecyl sulfate - polyacrylamide gel electrophoresis and Western - blot. Fluorescence activated cell sorting (FACS) antibody was used to detect PHMA02 in K562 and K562 / A02 cells Combined with activity, 125I-labeled antibody was modified by chloramine T method. The tail vein was injected into nude mice, SPECT scans were performed at different time points. Results The purity of PHMA02 was higher than that of P-gp +, and the positive rate of PMA-K562 / A02 cells was 94.47%. After 125I-labeled PHMA02 was injected into the P-gp + Radioactive enrichment occurred and was further enhanced over the next few days. Conclusion The imaging differences of 125I-PHMA02 antibody in P-gp + (K562 / A02) group and P-gp- (K562) control group can clearly differentiate P-gp + tumor tissue.