Generation and characterization of a monoclonal antibody against human BCL6 for immunohistochemical diagnosis

Abstract

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.

Detection of αB-Conotoxin VxXXIVA (αB-CTX) by ic-ELISA Based on an Epitope-Specific Monoclonal Antibody

“…To achieve high expression and purification of αB-CTX in E. coli BL21 (DE3), the DNA fragment encoding mature αB-CTX peptide was synthesized by Shanghai Biotech Co., Ltd. (Shanghai, China) after codon optimization [ 29 ]. The resulting DNA fragment was inserted into the plasmids pET32a and pBD-mbp for the construction of recombinant expression vectors, and the target proteins were expressed in E. coli BL21 (DE3) by IPTG induction (a final concentration of 1.0 mM) when the cultures reached an OD 600 of 1.0 [ 30 ].…”

A sensitive and rapid method of lead detection using nanoparticle technology based on monoclonal antibody

“…with 100 μg Pb-KLH. Three days later, splenocytes were collected and fused with SP2/0 cells at a ratio of 10:1 with PEG1450 ( Zhang et al, 2018 ; Jia et al, 2019 ). The fused cells were seeded into 96-well microplates and cultured in RPMI 1640 with a 20% FBS/HAT medium ( Zhang et al, 2018 ).…”

Evaluation of Apoptosis-Associated Speck-Like Protein (Asc) and B-Cell Lymphoma Protein (Bcl-6) in Relation to Peripheral Blood Biomarkers in Relapsing-Remitting Multiple Sclerosis

“…The concentrations of both ASC and BCL-6 (pg/ml) were determined using commercial ELISA kits according to the manufacturer's instructions (Inova Biotech company) [19,20].…”