Preparation and Characterization of a Monoclonal Antibody Against Human B Lymphocyte Stimulator

Sun, Jian; Li, Yan; Sun, Yingxun; Yu, Ming; Feng, Jie; Shen, Beifen

doi:10.1089/hyb.2006.25.80

Cited by 2 publications

(2 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Under the action of EDC, the introduced −COOH was coupled with the −NH 2 of the carrier protein BSA in the form of a monoamide bond (−CONH−) to synthesise the artificial antigen DON-BSA. For specific operation steps, refer to the EDC method described by Sun Zhen et al [ 31 ], with improvements. The detection antigen DON-OVA was prepared by the same method.…”

Section: Methodsmentioning

confidence: 99%

Comparison of Synthetic Methods and Identification of Several Artificial Antigens of Deoxynivalenol

Han

Jiang

et al. 2023

Molecules

View full text Add to dashboard Cite

The purpose of this experiment was to study the design and modification of hapten molecules and artificial antigen molecules of deoxynivalenol (DON), and to compare the preparation and identification methods of four artificial antigens. According to the characteristics of the molecular structure of DON, four artificial antigen coupling methods were designed—namely, N,N′-carbonyldiimidazole (CDI), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), isobutyl chloroformate (IBCF), and N-hydroxysuccinimide (NHS)—to prepare artificial antigens and detection antigens. Through ultraviolet (UV), infrared (IR), and SDS–polyacrylamide gel electrophoresis (SDS–PAGE), along with other physical and chemical identification methods and animal immunisation, the best artificial antigen coupling method was screened. The results showed that the CDI method achieved the best effect among the synthesis methods. The titre of anti-DON polyclonal antibody (pAb) produced by animal immunisation reached 1: (6.4 × 103). The half inhibitory concentration (IC50) was 47.75 ng/mL, the cross-reaction rate with 3-acetyldeoxynivalenol (3-AcDON) was slightly higher at 35.3%, and there was no cross-reaction with other compounds; therefore, four artificial antigens were successfully prepared by using the molecular structure of DON. Through identification, the CDI method was screened as the best artificial antigen synthesis method, with the highest DON pAb titre, the best sensitivity, and the strongest specificity. This will lay a solid antigenic foundation for the preparation of better anti-DON monoclonal antibodies (mAbs) in the future.

show abstract

Section: Methodsmentioning

confidence: 99%

Comparison of Synthetic Methods and Identification of Several Artificial Antigens of Deoxynivalenol

Han

Jiang

et al. 2023

Molecules

View full text Add to dashboard Cite

show abstract

“…Recombinant human soluble BLyS (rBLyS) and anti-human BLyS MAb B7, (18) and recombinant human TRAIL (rTRAIL) were prepared in our laboratory. Human lymphoma cell lines (U937, Jurkat, Daudi, and Raji) and MM cell lines (XG-7 and SKO-007) were stored in our laboratory.…”

Section: Methodsmentioning

confidence: 99%

Analysis of an Anti–B Lymphocyte Stimulator Monoclonal Antibody B7 and Its Binding Activity to Myeloma and Lymphoma Cell Lines

Sun

Li²,

Yu³

et al. 2006

Hybridoma

Self Cite

View full text Add to dashboard Cite

Although there is convincing evidence of a link between B lymphocyte stimulator (BLyS) and the proliferation and survival of malignant B cells, previous observations about BLyS expression on B lymphoma cells were contradictory. In this study, BLyS expression on human lymphoma and myeloma cell lines was evaluated by Fluorescence-Activated Cell Sorter (FACS). First, specificity of a monoclonal antibody (MAb) against BLyS, was analyzed. The results showed that MAb B7 was immunoglobulin G(3) (IgG(3)) and recognized recombinant human BLyS specifically. In addition, MAb B7 bound to the histiocyte lymphoma cell line U937 in dosage-dependent manner, but not the T lymphoma cell line Jurkat, suggesting that the cellular binding of MAb B7 was specific. Using this MAb, BLyS expression on two multiple myeloma cell lines (XG-7 and SKO-007) and two Burkitt's lymphoma cell lines (Daudi and Raji) was evaluated. MAb B7 did bind to XG- 7 and SKO-007 (66.84% and 79.38% positive cells, respectively). But MAb B7 did not bind Burkitt's lymphoma cell lines, Daudi and Raji (4.09% and 3.02% positive cells, respectively). It will be interesting to further analyze the expression of BLyS on B malignant cells of multiple myeloma and lymphoma patients, and to evaluate the correlation of its expression and patient prognosis.

show abstract

Preparation and Characterization of a Monoclonal Antibody Against Human B Lymphocyte Stimulator

Cited by 2 publications

References 27 publications

Comparison of Synthetic Methods and Identification of Several Artificial Antigens of Deoxynivalenol

Comparison of Synthetic Methods and Identification of Several Artificial Antigens of Deoxynivalenol

Analysis of an Anti–B Lymphocyte Stimulator Monoclonal Antibody B7 and Its Binding Activity to Myeloma and Lymphoma Cell Lines

Contact Info

Product

Resources

About