期刊论文

Preparation and identification of rabbit polyclonal antibody against BCR-ABL b3a2 fusion protein

中文

细胞与分子免疫学杂志

1007-8738

2021

37

08

746-751

10.13423/j.cnki.cjcmi.009214

湖南省自然科学基金（2017JJ2004）； 第九批博士后科学基金特别资助项目（2016T90765）； 湖南省卫计委课题（C20180166）；

本校为第一机构

目的制备兔抗断裂点集簇区-Abelson鼠白血病病毒癌基因（BCR-ABL）b3a2位点的多克隆抗体并鉴定其特异性。方法针对BCR-ABL融合蛋白b3a2型的特异性抗原序列,设计并合成纯度高于90%的多肽。一方面通过透孔螺血蓝蛋白（KLH）偶联融合多肽用于免疫新西兰兔,多次加强免疫得到抗血清;另一方面通过仅偶联单独BCR或者ABL肽段对抗血清进行二次纯化,吸附并去除未结合融合表位的抗血清以达到进一步纯化抗体的效果。最终得到的抗血清用于ELISA及Western blot法检测。结果免疫前兔血清背景均低,免疫后抗血清效价达到1∶32 000,满足实验要求。抗原亲和纯化所得抗体纯度及效价高,Western blot法检测表明抗血清能...

Objective To prepare and identify rabbit anti-breakpoint cluster region-Abelson leukemia virus oncogene (BCR-ABL) b3a2 subtype polyclonal antibody. Methods A peptide containing the fusion sequence of the b3a2 subtype BCR-ABL fusion protein was designed and synthesized with the purity higher than 90%. The fusion polypeptide was coupled to Keyhole Limpet hemocyanin (KLH) and used to immune New Zealand rabbits. Antiserum was purified after multiple immunizations, in addition to using the b3a2 subtype fusion polypeptide for affinity purification. Peptides harboring only BCR or c-ABL amino acid seq...