CAJ | 학술논문

目的制备一种高灵敏、高特异的免疫磁性微球(IMMS),为其在肿瘤早期诊断方面的应用提供借鉴.方法分别采用N-羟基琥珀酰亚胺3-(2-吡啶二硫代基丙酸酯)(SPDP)和1-乙基-3-(3-二甲基胺基丙基)碳化二亚胺盐酸盐(EDC)作为交联剂,将鼠抗人免疫球蛋白G(IgG)偶联于磁性微球上.采用激光共聚焦检测和125Ⅰ标记抗体考察偶联效果.比较交联剂、磁球微球上功能团、间隔臂、偶联条件及抗体用量等因素对偶联效果的影响.分别对不同浓度人源IgG进行吸附,确定最低检测值.结果实验结果表明间隔臂对偶联效果影响不大.两种交联剂均可将抗体联接于磁性微球上,EDC为交联剂,工艺更为简便,室温条件下抗体结合效果优于4℃条件下.本实验所制备的IMMS对人源IgG的最低检测值为50pg/mL.结论本实验以人源IgG为肿瘤标记模型,成功地制备了一种高灵敏、高特异免疫磁性微球,制备简便,成本低廉,为肿瘤早期诊断提供了借鉴.
목적 제비일충고령민、고특이적면역자성미구(IMMS),위기재종류조기진단방면적응용제공차감.방법분별채용N-간기호박선아알3-(2-필정이류대기병산지)(SPDP)화1-을기-3-(3-이갑기알기병기)탄화이아알염산염(EDC)작위교련제,장서항인면역구단백G(IgG)우련우자성미구상.채용격광공취초검측화125Ⅰ표기항체고찰우련효과.비교교련제、자구미구상공능단、간격비、우련조건급항체용량등인소대우련효과적영향.분별대불동농도인원IgG진행흡부,학정최저검측치.결과 실험결과표명간격비대우련효과영향불대.량충교련제균가장항체련접우자성미구상,EDC위교련제,공예경위간편,실온조건하항체결합효과우우4℃조건하.본실험소제비적IMMS대인원IgG적최저검측치위50pg/mL.결론 본실험이인원IgG위종류표기모형,성공지제비료일충고령민、고특이면역자성미구,제비간편,성본저렴,위종류조기진단제공료차감.
Objective To develop a novel immunomagnetic microsphere(IMMS) with high sensitivity and specificity, providing reference for early diagnosis of tumors. Methods The murine anti-human IgG was covalently bound on the MMS by using SPDP or EDC as coupling agent. The coupling effect was evaluated by laser scanning confocal microscope (LSCM), and the antibody and antigen bound on the MMS were quantitated by radioimmunoassay. Several influential factors were investigated, including coupling agents, functional groups on the MMS, space arms, reaction time and temperature, and the amount of antibody, etc. The lowest detectable concentration of human IgG was determined. Results Using either SPDP or EDC as coupling agents, the antibody was effective bound on the MMS. The spacer length had no significant influence on the coupling efficiency. Even though both EDC and SPDP were able to link the antibody to the MMS, EDC seems to be a more efficient coupling agent since the coupling process was simpler and faster. More antibodies were bound on the MMS when the cross-linking reaction was conducted at 25℃ for 3 h than that of at 4℃ for 18 h. The detection limit for human IgG was 50 pg/ml. Conclusion In this paper, a IMMS with high sensitivity and specificity was successfully synthesized using human IgG as an antigen model. Given the cheapness and high specificity of this method, it might be a promising approach for detecting proteins in trace amount.