国际放射医学核医学杂志
國際放射醫學覈醫學雜誌
국제방사의학핵의학잡지
INTERNATIONAL JOURNAL OF RADIATION MEDICINE AND NUCLEAR MEDICINE
2010年
2期
107-109
,共3页
洪智慧%周小林%石怡珍%刘增礼
洪智慧%週小林%石怡珍%劉增禮
홍지혜%주소림%석이진%류증례
碘放射性同位素%胃泌素释放钛%同位素标记%免疫活性%单链抗体
碘放射性同位素%胃泌素釋放鈦%同位素標記%免疫活性%單鏈抗體
전방사성동위소%위비소석방태%동위소표기%면역활성%단련항체
Iodine radioisotopes%Gastrin-releasing peptide%Isotope labeling%Immunocompetence1%Single chain antibody
目的 研究抗胃泌素释放前体(ProGRP(31-98))单链抗体(scFv)的131I标记方法,并对其标记后的稳定性和免疫活性进行分析.方法 采用氯胺-T法碘化标记制备131I-anti-ProGRP(31-98)scFv,凝胶柱层析法分离纯化标记产物,利用纸层析法测定标记物的标记率、放化纯和稳定性,通过细胞结合试验测定131I-anti-ProGRP(31-98)scFv的免疫活性.结果 131I-anti-ProGRP(31-98)标记率为93.35%.标记产物纯化后即刻放化纯为98.49%,在37℃水浴箱中放置24 h后放化纯为94.59%,48h后测定仍大于90%,与正常人血清充分混合24 h后放化纯为85.16%,48 h测定大于80%.131I-anti-ProGRP(31-98) scFv对人小细胞肺癌NCI-H446和肺腺癌A549细胞株的免疫结合率分别为85.36%和21.02%.结论 131I-anti-ProGRP(31-98)scFv的标记率高,且有良好的稳定性和免疫活性,有望成为高表达ProGRP的恶性肿瘤放射免疫显像及治疗药物,值得进一步深入研究.
目的 研究抗胃泌素釋放前體(ProGRP(31-98))單鏈抗體(scFv)的131I標記方法,併對其標記後的穩定性和免疫活性進行分析.方法 採用氯胺-T法碘化標記製備131I-anti-ProGRP(31-98)scFv,凝膠柱層析法分離純化標記產物,利用紙層析法測定標記物的標記率、放化純和穩定性,通過細胞結閤試驗測定131I-anti-ProGRP(31-98)scFv的免疫活性.結果 131I-anti-ProGRP(31-98)標記率為93.35%.標記產物純化後即刻放化純為98.49%,在37℃水浴箱中放置24 h後放化純為94.59%,48h後測定仍大于90%,與正常人血清充分混閤24 h後放化純為85.16%,48 h測定大于80%.131I-anti-ProGRP(31-98) scFv對人小細胞肺癌NCI-H446和肺腺癌A549細胞株的免疫結閤率分彆為85.36%和21.02%.結論 131I-anti-ProGRP(31-98)scFv的標記率高,且有良好的穩定性和免疫活性,有望成為高錶達ProGRP的噁性腫瘤放射免疫顯像及治療藥物,值得進一步深入研究.
목적 연구항위비소석방전체(ProGRP(31-98))단련항체(scFv)적131I표기방법,병대기표기후적은정성화면역활성진행분석.방법 채용록알-T법전화표기제비131I-anti-ProGRP(31-98)scFv,응효주층석법분리순화표기산물,이용지층석법측정표기물적표기솔、방화순화은정성,통과세포결합시험측정131I-anti-ProGRP(31-98)scFv적면역활성.결과 131I-anti-ProGRP(31-98)표기솔위93.35%.표기산물순화후즉각방화순위98.49%,재37℃수욕상중방치24 h후방화순위94.59%,48h후측정잉대우90%,여정상인혈청충분혼합24 h후방화순위85.16%,48 h측정대우80%.131I-anti-ProGRP(31-98) scFv대인소세포폐암NCI-H446화폐선암A549세포주적면역결합솔분별위85.36%화21.02%.결론 131I-anti-ProGRP(31-98)scFv적표기솔고,차유량호적은정성화면역활성,유망성위고표체ProGRP적악성종류방사면역현상급치료약물,치득진일보심입연구.
Objective To study the 131I labeling methods, stability and immunological activity of antiprogastrin-releasing peptide(31-98) single-chain antibody (anti-ProGRP(31-98) scFv). Methods The anti-ProGRP(31-98)scFv was labeled by Chloramine-T method usng 131I, and purified by gel column separation method.The labeling efficiency, radiochemical purity and stability were estimated by using paper chromatography.Immunological activity was detected with cell conjugation assay. Results The labeling efficiency and radiochemical purity of 131I-anti-ProGRP(31-98) scFv were 93.35% and 98.49%, respectively. The radiochemical purity was 94.59% after incubation in water bath at 37℃ for 24h, and still maintained above 90% for 48 h.After incubation with healthy human serum for 48 h, the radiochemical purity was still above 80%. The immunobinding ratios of 131I-anti-ProGRP(31-98)scFv for NCI-H446 cells and A549 cells were 85.36% and 21.02%, respectively. Conclusions 131I-anti-ProGRP(31-98)scFv not only has high labeling efficiency and radiochemical purity, but also has good stability and keeps good immunological activity. Therefore, it could be a promising agent for radioimmunoimaging and radioimmunotherapy for the malignant tumors which highly express ProGRP.
