[Construction and expression of human 4-1BBL/anti-CD20 fusion protein].
OBJECTIVE
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.


CONCLUSIONS
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.
Liu
Rong
R
Institute of Hematology & Hospital of Blood Disease, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin, China.
Jiang
Wen-guo
WG
Liu
Fang
F
Zhang
Yan-jun
YJ
Fan
Dong-mei
DM
Yang
Ming
M
Xiong
Dong-sheng
DS
Yang
Chun-zheng
CZ
D053330:4-1BB Ligand / Q000235:genetics* / Q000378:metabolism; D000906:Antibodies / Q000235:genetics* / Q000378:metabolism; D018951:Antigens, CD20 / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D053330:4-1BB Ligand; D000906:Antibodies; D018951:Antigens, CD20; D011993:Recombinant Fusion Proteins