[Construction and expression of a functional chimeric murine-human antibody directing against human cross-linked fibrin].
OBJECTIVE
In order to reduce the immunogenicity of the murine antifibrin monoclonal antibody (MAb) SZ-63 to human beings.


METHODS
First of all, the cDNA sequences encoding the constant regions of the murine kappa light chain and gamma 1 heavy chain in the SZ-63 MAb were substituted with the corresponding human genomic sequences. Then the constructed MAb was cloned separately into two selectable expression vectors, and the latter were transfected into mouse myeloma cells (SP2/0) one by one.


RESULTS
ELISA, Western blot and competition experiment results showed that there was 0.8 approximately 1.0 mg/L of chimeric 63 IgG in the conditioned medium of selected cell lines, and the expressed IgG had higher antifibrin D-Dimer capability than that of the original MAb.


CONCLUSIONS
The expressed chimeric antibody can be used as a targeting agent for thrombus imaging and treatment.
Liu
Y
Y
Thrombosis and Hemostasis Research Unit, Jiangsu Institute of Hematology, Suzhou Medical College, Suzhou 215007.
Xia
L
L
Fu
J
J
Ruan
C
C
Wan
H
H
Li
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology*; D001667:Binding, Competitive; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005337:Fibrin / Q000737:chemistry* / Q000276:immunology*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D005337:Fibrin