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一种靶向B细胞淋巴瘤B细胞活化因子的新型VHH抗体。

A novel VHH antibody targeting the B cell-activating factor for B-cell lymphoma.

作者信息

Wu Wen, Li Shenghua, Zhang Weijing, Sun Jian, Ren Guangda, Dong Quanchao

机构信息

Department of Lymphoma, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, China.

Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin Shengfa NabioTech Co., Ltd., Tianjin 300457, China.

出版信息

Int J Mol Sci. 2014 May 28;15(6):9481-96. doi: 10.3390/ijms15069481.

DOI:10.3390/ijms15069481
PMID:24879522
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4100105/
Abstract

OBJECTIVE

To construct an immune alpaca phage display library, in order to obtain a single domain anti-BAFF (B cell-activating factor) antibody.

METHODS

Using phage display technology, we constructed an immune alpaca phage display library, selected anti-BAFF single domain antibodies (sdAbs), cloned three anti-BAFF single-domain antibody genes into expression vector pSJF2, and expressed them efficiently in Escherichia coli. The affinity of different anti-BAFF sdAbs were measured by Bio layer interferometry. The in vitro biological function of three sdAbs was investigated by cell counting kit-8 (CCK-8) assay and a competitive enzyme-linked immunosorbent assay (ELISA).

RESULTS

We obtained three anti-BAFF single domain antibodies (anti-BAFF64, anti-BAFF52 and anti-BAFFG3), which were produced in high yield in Escherichia coli and inhibited tumor cell proliferation in vitro.

CONCLUSION

The selected anti-BAFF antibodies could be candidates for B-cell lymphoma therapies.

摘要

目的

构建免疫羊驼噬菌体展示文库，以获得单域抗B细胞激活因子（BAFF）抗体。

方法

利用噬菌体展示技术构建免疫羊驼噬菌体展示文库，筛选抗BAFF单域抗体（sdAbs），将3个抗BAFF单域抗体基因克隆到表达载体pSJF2中，并在大肠杆菌中高效表达。采用生物膜干涉技术测定不同抗BAFF sdAbs的亲和力。通过细胞计数试剂盒-8（CCK-8）法和竞争性酶联免疫吸附测定（ELISA）研究3种sdAbs的体外生物学功能。

结果

获得3个抗BAFF单域抗体（抗BAFF64、抗BAFF52和抗BAFFG3），它们在大肠杆菌中高产表达，并在体外抑制肿瘤细胞增殖。

结论

筛选出的抗BAFF抗体可作为B细胞淋巴瘤治疗的候选药物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/857af41f280a/ijms-15-09481-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/a62dc7a0c8f0/ijms-15-09481-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/8cb83ecc8f67/ijms-15-09481-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/77a9f71f2772/ijms-15-09481-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/2c3ed0d5dcd0/ijms-15-09481-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/857af41f280a/ijms-15-09481-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/a62dc7a0c8f0/ijms-15-09481-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/8cb83ecc8f67/ijms-15-09481-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/77a9f71f2772/ijms-15-09481-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/2c3ed0d5dcd0/ijms-15-09481-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a317/4100105/857af41f280a/ijms-15-09481-g005.jpg

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