Open AccessGeneration and characterization of a monoclonal antibody against human BCL6 for immunohistochemical diagnosis
Author(s) -
Kunzhi Jia,
Danping Zhang,
Yanghai Wang,
Yaju Liu,
Xiangpei Kong,
Qinghai Yang,
Huiling Chen,
Cheng-Jie Xie,
Shihua Wang
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0216470
Subject(s) - immunohistochemistry , monoclonal antibody , bcl6 , antibody , monoclonal , biology , pathology , microbiology and biotechnology , virology , medicine , immunology , b cell , germinal center
Background Human B-cell lymphoma 6 (BCL6) gene, usually coding protein of 706 amino acids, is closely associated with large B cell lymphoma. Researches showed that protein mutation or change of expression levels usually happened in the mounting non-hodgkin lymphoma (NHL). Thus BCL6 is considered to be involved in germinal center (GC)-derived lymphoma. Results The BCL6 1-350 gene codons were optimized for prokaryotic system. After expression of BCL6 1-350 in E . coli , the BCL6 1-350 protein was purified with Ni column. Then the BCL6 1-350 protein, mixing with QuickAntibody-Mouse5W adjuvant, was injected into Balb/c mice. After immunization and cell fusion, a stable cell line named 1E6A4, which can secrete anti-BCL6 antibody, was obtained. The isotype of 1E6A4 mAb was determined as IgG 2a , and the affinity constant reached 5.12×10 10 L/mol. Furthermore, the specificity of the mAb was determined with ELISA, western blot and immunohistochemistry. Results indicated that the 1E6A4 mAb was able to detect BCL6 specifically and sensitively. Conclusions BCL6 1-350 antigen has been successfully generated with an effective and feasible method, and a highly specific antibody named 1E6A4 against BCL6 has been screened and characterized in this study, which was valuable in clinical diagnosis.