抗人乳腺癌单克隆抗体偶联米托蒽醌白蛋白纳米球的初步研究

AIM To improve the treatment efficacy of anti tumor drug mitoxantrone, the conjugation of mitoxantrone loaded nanospheres and anti C erbB 2 monoclonal antibodies were prepared. METHODS Mitoxantrone loaded nanospheres were prepared with emulsion heating solidification technique. A heterobifunctional reagent, N succinimidyl 3 (2 pyridyldithio) propionate (SPDP), was used as the crosslinker of mitoxantrone loaded nanospheres and anti C erbB 2 monoclonal antibodies; pharmaceutical properties of immunonanocapsuls were studied; the conjugates of nanospheres and monoclonal antibodies was confirmed with immunological methods such as slide agglutination test, fluorescent immunossay and rosset formation test, fluorescent staining and scanning electron microscope. RESULTS Mitoxantrone loaded nanospheres were spherical, with smooth surface and median diameter of 0 665 micron. When stored at 3-5, 20-25 and 37℃, RH 75% for three months, the appearance, morphology, size distribution, drug loading and in vitro release characteristics showed no significant change and the stability was satisfactory. The size analysis demonstrated that there was no obvious increase in the particle size of nanoparticles after conjugation. Immunological tests indicate highly selective binding of antibody targeted nanospheres to C erbB 2 overexpressing cells SK BR 3. CONCLUSION The conjugation of mitoxantrone loaded nanospheres and anti C erbB 2 monoclonal antibodies can keep the activity of anti C erbB 2 and increase the therapeutic efficacy of anti mammary cancer drugs.

抗乳腺癌单抗CDI315B可变区基因的序列分析

目的 :克隆抗乳腺癌 CDI315 B单抗可变区基因 ,并对其进行序列分析。方法 :利用前导肽序列设计引物 ,采用 RT-PCR技术从分泌抗人乳腺癌单克隆抗体的杂交瘤细胞株 CDI315 B分离克隆了抗体轻重链可变区基因 ,用 Sanger双脱氧末端终止法测定扩增的 CDI315 B单抗 VK 和 VH 基因的序列 ,对其进行序列分析。结果 :和国际标准的小鼠 Kabat分类体系进行对比分析 ,CDI315 B单抗的 VK属于第 4或第 5组的第 6亚组 (VI) ;CDI315 B单抗的 VH 则属于第 2族 (VH2 )。结论 :抗乳腺癌 CDI315 B单抗可变区基因的克隆及序列分析为构建人鼠嵌合轻链和人鼠嵌合重链打下基础。