抗P-gp抗体PHMA02在裸鼠体内的影像分布研究

Investigation of photographic distribution of anti-P-gp antibody PHMA02 in nude mice

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 investigate the photographic distribution of anti-P-gp monoclonal antibody PHMA02 in nude mice transplanted with K562/A02 cell tumor. Methods PHMA02 were prepared by hybridoma technique, sodium dodecyl sulfate-polyacrylamide gel electrophorsis and Western-blot were used to analyzed its purity, fluorescence-activated cell sorting（FACS） was also used to determine its binding with K562 and K562/A02 cells. Nude mice were injected i.v with ^125I labeled PHMA02, and were scanned on different time by SPECT. Results The purity and binding affinity of PHMA02 were both satisfactory. On the second day from injection, radioactivity appeared in all tumor regions of the P-gp＋（K562/A02） group nude mice, and intensified in the following days. Concludsion The photographic difference of PHMA02 distribution between P-gp＋（K562/A02） and P-gp-（K562） group could specifically distinguish the P-gp^＋ tumor tissue.