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Updated: Jul 4, 2026
Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
[Construction and expression of human 4-1BBL/anti-CD20 fusion protein]
Rong Liu1, Wen-guo Jiang, Fang Liu
1Institute of Hematology & Hospital of Blood Disease, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin, China.
Aim:
To construct and express human 4-1BBL/anti-CD20 bispecific fusion protein and identify its biological activity.
Methods:
PCR and overlapping PCR were used to construct human 4-1BBL/anti-CD20 bispecific fusion protein. DNA sequencing was performed by the terminus of the fusion protein nucleotide.The product was purified by affinity chromatography and analyzed by Western blot and its antigen-binding activity was examined by FACS.
Results:
The data of DNA sequence showed that human 4-1BBL/anti-CD20 bispecific fusion protein was correct. The fusion protein was recovered in high yield (up to 4 mg/L) after E-tag purification and predominantly(90%) as a dimer. The fusion protein could bind to Raji cells(CD20(+)) and A549 cells(4-1BB(+)), respectively.
Conclusion:
The human 4-1BBL/anti-CD20 bispecific fusion protein with high level expression was successfully obtained and could bind to Raji ceIls and A549 cells.
