江西科学
江西科學
강서과학
JIANGXI SCIENCE
2012年
5期
590-593
,共4页
母波%赵春艳%梁素华%章欢%申跃武%陈保峰%杨俊宝
母波%趙春豔%樑素華%章歡%申躍武%陳保峰%楊俊寶
모파%조춘염%량소화%장환%신약무%진보봉%양준보
多克隆抗体%多发性骨髓瘤%制备
多剋隆抗體%多髮性骨髓瘤%製備
다극륭항체%다발성골수류%제비
Polyclonal antibody%Multiple myeloma%Preparation
制备人多发性骨髓瘤全细胞兔多克隆抗体。用人多发性骨髓瘤细胞系ARH-77全细胞和福氏完全佐剂通过背部皮内多点注射首次免疫新西兰大白兔,第14 d用ARH-77全细胞和福氏不完全佐剂同样剂量加强免疫,第28 d和38 d时再次免疫。第45 d颈动脉采血80 mL,4℃静置过夜,收集血清。然后采用亲和层析系统纯化血清中的多克隆抗体。用酶联免疫吸附试验(ELISA)检测纯化多克隆抗体的效价;免疫印迹法和荧光免疫细胞化学检测纯化多克隆抗体的特异性。结果:ELISA检测多克隆抗体滴度为1∶20 000,免疫印迹检测结果显示该抗体具有较高特异性,荧光免疫细胞化学检测多克隆抗体能够有效的结合细胞表面抗原。实验获得了高效价的特异性的兔抗人骨髓瘤细胞多克隆抗体,为进一步研究多发性骨髓瘤诊断和治疗奠定了基础。
製備人多髮性骨髓瘤全細胞兔多剋隆抗體。用人多髮性骨髓瘤細胞繫ARH-77全細胞和福氏完全佐劑通過揹部皮內多點註射首次免疫新西蘭大白兔,第14 d用ARH-77全細胞和福氏不完全佐劑同樣劑量加彊免疫,第28 d和38 d時再次免疫。第45 d頸動脈採血80 mL,4℃靜置過夜,收集血清。然後採用親和層析繫統純化血清中的多剋隆抗體。用酶聯免疫吸附試驗(ELISA)檢測純化多剋隆抗體的效價;免疫印跡法和熒光免疫細胞化學檢測純化多剋隆抗體的特異性。結果:ELISA檢測多剋隆抗體滴度為1∶20 000,免疫印跡檢測結果顯示該抗體具有較高特異性,熒光免疫細胞化學檢測多剋隆抗體能夠有效的結閤細胞錶麵抗原。實驗穫得瞭高效價的特異性的兔抗人骨髓瘤細胞多剋隆抗體,為進一步研究多髮性骨髓瘤診斷和治療奠定瞭基礎。
제비인다발성골수류전세포토다극륭항체。용인다발성골수류세포계ARH-77전세포화복씨완전좌제통과배부피내다점주사수차면역신서란대백토,제14 d용ARH-77전세포화복씨불완전좌제동양제량가강면역,제28 d화38 d시재차면역。제45 d경동맥채혈80 mL,4℃정치과야,수집혈청。연후채용친화층석계통순화혈청중적다극륭항체。용매련면역흡부시험(ELISA)검측순화다극륭항체적효개;면역인적법화형광면역세포화학검측순화다극륭항체적특이성。결과:ELISA검측다극륭항체적도위1∶20 000,면역인적검측결과현시해항체구유교고특이성,형광면역세포화학검측다극륭항체능구유효적결합세포표면항원。실험획득료고효개적특이성적토항인골수류세포다극륭항체,위진일보연구다발성골수류진단화치료전정료기출。
To prepare polyclonal antibody of rabbit anti-human multiple myeloma. Method:The New Zealand white rabbits were immunized with human multiple myeloma ARH-77 cell line at the back skin intradermal injection for the first time, he primary inoculation was with Freund's complete adju- vant (Sigma) ,follow by three boosts with Freund's incomplete adjuvant (Sigma) on the 14,28 and 38day. The serum was extracted after 4 ℃ settlement overnight. Polyclonal antibodies were isolated using affinity chromatography system. Purified polyclonal antibodies was analyzed by ELISA, Westem blot and Immunofluorescence stain. Results:polyclonal antibody titer was up to 1 : 20 000 by ELISA. Western blot showed that the antibody had a high specificity. Immunofluorescence stain showed Poly- clonal antibodies could specificitily binding with ARH-77 cell surface proteins. Conclusion:We pro- duced polyclonal antibodies of rabbit anti-human multiple myeloma and it have high titer and speci- ficity, to providing a new method for diagnosis and therapy of multiple myeloma.