recombinant human antibody production problem - (Aug/08/2008 )
Dear all,
I have used a mammelian expression vector pCAGGS in order to express the recombinant antibodies.
The vector contains a chicken beta actin promotor with CAG enhancer followed by secretion leader sequence, mouse variable heavy chain, human gamma1 constant chain, internal ribosome entry site, (IRES), mouse variable light chain and human kappa chain respectively. My indicator plasmid contains green fluorescent protein (GFP) gene in pCAGGS. When I transfect the indicator plasmid, it shows efficient expression, while my construct which is supposed to produce antibody does not show it in the supernatant where it should be. Since I am doing this thing for the first time, I do not know what is wrong. Thank you in advance for your suggestions and comments.
I have used a mammelian expression vector pCAGGS in order to express the recombinant antibodies.
The vector contains a chicken beta actin promotor with CAG enhancer followed by secretion leader sequence, mouse variable heavy chain, human gamma1 constant chain, internal ribosome entry site, (IRES), mouse variable light chain and human kappa chain respectively. My indicator plasmid contains green fluorescent protein (GFP) gene in pCAGGS. When I transfect the indicator plasmid, it shows efficient expression, while my construct which is supposed to produce antibody does not show it in the supernatant where it should be. Since I am doing this thing for the first time, I do not know what is wrong. Thank you in advance for your suggestions and comments.
What type of mammalian cells are you using?
Its 293T.....any clue please?